Conjugates of toll-like receptor 7 or 8 agonists with temporarily inactivated activation sites and functional drugs, and uses thereof

Abstract

To provide a drug composition utilizing synergistic effect of a functional drug and toll-like receptor 7 or 8 agonist substance.SOLUTION: The present invention relates to toll-like receptor 7 or 8 agonist and a drug conjugate substance in which a functional drug and toll-like receptor 7 or 8 agonist are linked by a chemical bond containing a cleavable site, and uses thereof. Side effects of toll-like receptor 7 or 8 agonist can be reduced by using the property of being temporarily inactivated, while utilizing the synergistic effect of the functional drug and toll-like receptor 7 or 8 agonist substance. In particular, the chemical bond is designed to inhibit an immune activation function of toll-like receptor 7 or 8 agonist, and then restore the immune activation function at an injection site, or within a tumor microenvironment or in a target immune cells, thereby showing kinetic properties that can regulate immune activation efficacy over time, and enabling the minimization of non-specific immune response induction.SELECTED DRAWING: Figure 22

Description

[Technical field]

[0001] The present invention relates to a binding agent in which a functional drug is bound to the activation site of a toll-like receptor 7 or 8 agonist. More specifically, the active site of Toll-like receptor 7 or 8 agonists. Functional drugs bind to the Toll-like receptor 7 or 8 agonist site, temporarily inhibiting the function of the receptor. is inhibited at a certain time interval and then functions dynamically. is kinetically recovered. Regarding its uses, etc. [Background technology]

[0002] The body's innate immune response is a key player in the prevention and treatment of a variety of infectious, cancer, metabolic, and rare diseases. However, patients with certain diseases may not be able to fully understand the In many cases, normal immune response functions such as the immune system are impaired. Strengthening the body's weakened immune response as a possible method for preventing, improving and treating diseases. or reactivate immune responses that have been suppressed or halted by specific signals. Various functional drugs have been developed. Generally, immune responses are caused by activated immune cells acting against foreign substances. and a series of reactions against endogenous substances, i.e., antigens. When microorganisms, including bacteria and viruses, or foreign substances enter the body, immune cells It recognizes this and becomes activated, secreting factors such as cytokines to induce an inflammatory response. Recently, there has been active research into the mechanisms of innate immune responses that act nonspecifically in the early stages of infection. Among these, Toll-like receptor (T LR) is a receptor that can recognize pathogens at the early stage of inflammation and binds to plasma membrane components and nucleic acids of pathogens. It is known that the immune system can recognize structures and induce immune responses, and this can be used to Research on various Toll-like receptor ligands for activating This is being carried out regularly.

[0003] Among various Toll-like receptor ligands, Toll-like receptor 7 or 8 agonists (agonis t)-based substances are used as immunoadjuvants to induce cell-mediated immune responses and Imidazoquinoline-based agonists t), hydroxyadenine-based agonists (8-hydroxyadenine-based agonist, pteridone-based agonist st), 2-aminopyrimidine-based agonist, benzoazepine-based agonist, thiaoxoguanosine agonist (7-thia-8-oxogua nosine-based agonists are known (US Patent Publication 201 Such Toll-like receptor 7 or 8 agonists are known to mediate endosome activation. It is an agonist of toll-like receptors 7 and 8 in the body, and has been shown to activate not only humoral immunity but also cellular immunity. Recently, Toll-like receptor 7 or 8 agonists have been reported to effectively induce Beyond its adjuvant function, it has been shown that it exerts an immunosuppressive effect at the injection site or in the tumor microenvironment. This leads to the suppression of immune-suppressing cells, such as MDSCs, Tregs, and M2 mac cells, which aid in cancer growth and metastasis. By inhibiting the function of rophage, etc., it enhances the immune anti-cancer effect. However, the multifunctional Toll-like receptor Due to their molecular structure, it is difficult for the 7th or 8th agonist drugs to disperse in an aqueous solution. It is only soluble in special organic solvents such as DMSO, methanol, etc., and is commonly used Since it is not soluble in organic solvents, it has limitations in producing immune-activating drugs in various dosage forms. Therefore, cream-type formulations containing a mixture of various surfactants (e.g., Ald Some research has been conducted to overcome these problems. In order to be effective, it is prepared in the form of a salt and can be dissolved in an aqueous solution. However, toll-like receptor 7 or 8 agonists produced in the form of salts were absorbed into the bloodstream in the body. This induces a systemic immune response in the blood vessels. By doing so, many side effects (e.g., cytokine storm) can be prevented. It is not easy to use because it induces various non-specific hypersensitivity immune reactions, etc. In addition, due to the problem of side effects, it is practically impossible to use it for treatment. To prevent this, a concentration less than the effective dose must be used. Some pharmaceutical companies are trying to overcome this problem by In addition, lipids that exhibit lipophilic properties can be introduced into the polymer chains, and the polymer chains can be directly chemically bonded to the polymer chains. Attempts have also been made to prevent direct absorption into the bloodstream by binding it to However, the Toll-like receptor 7 or 8 agonist produced by such a method Since the active site is still exposed to the outside, However, there is still a possibility that it may induce toxicity by inducing a non-specific immune response in is.

[0004] Thus, chemical conjugation of the activation site could allow immunotherapy of Toll-like receptor 7 or 8 agonists. The activation function is inhibited and then distributed to the injection site, the tumor microenvironment, or is designed to restore immune activation function in target immune cells. These can be engineered to exhibit kinetic properties that allow for modulation of immune activation efficacy over time. If this is possible, it may be possible to prevent nonspecific toxicity and immune response induction by Toll-like receptor 7 or 8 agonists. In addition, various functional drugs (antibodies, small molecule anticancer drugs, antigens, Cytokines, peptides, amino acids, small molecules, lipids, oligonucleotides, targets Toll-like receptor 7 or 8 agonist drugs are combined with functional drugs to bind toll-like receptor 7 or 8 agonist drugs. By utilizing the synergistic effects of these two molecules, Diverse applications of regulating cancer cell death, therapeutic immune cell activation, immune suppressor cells and the environment It is expected that it can be used widely in pharmaceutical compositions. Summary of the Invention [Problem to be solved by the invention]

[0005] The present invention has been made to solve the problems in the prior art as described above, Transient inhibition of immune stimulatory functions of Toll-like receptor 7 or 8 agonists n) and then specifically detect immune activity at the injection site, in the tumor microenvironment, or within target cells. Toll-like receptor 7 or 8 agonists designed to restore A conjugate of a functional drug and a toll-like receptor 7 or 8 agonist, and its use The purpose of this document is to provide a means for

[0006] However, the technical problem that the present invention aims to achieve is not limited to the problems mentioned above. Further problems not mentioned above and not mentioned can be solved by the following description, which is a conventional method in the art. Engineers can understand it clearly. [Means for solving the problem]

[0007] The present invention relates to a toll-like receptor 7 or 8 agonist and a functional drug. a conjugate of a functional drug and a Toll-like receptor, The active site of the receptor 7 or 8 agonist is linked to a cleavable linker ( The present invention provides a conjugate characterized in that the conjugate is bound to a cleavable linker. More particularly, the conjugate binds to the activation site of Toll-like receptor 7 or 8 agonists. ive site), i.e., exhibits biological activity of Toll-like receptor 7 or 8 agonists Toll-like receptor 7 or The 8-agonist is temporarily inactivated, and a functional drug is attached to the other end of the linker. It is a chemically bonded conjugate.

[0008] In one embodiment of the invention, the conjugate is a conjugate in which the functional drug acts primarily in an inactive form. Toll-like receptor 7 or 8 agonists in the tumor-like receptor 7 or 8 domain are targeted to the tumor at the injection site, the tumor microenvironment or target cells. The linker is cleaved within the kinetically activated site. It is characterized by the recovery of the primary site and acting secondarily.

[0009] In another embodiment of the invention, the cleavable linker is sensitive to enzymes, pH, redox potential, from the group consisting of redox potential, temperature, ultrasound, magnetism, and light source. The chemical bond at the binding site is cleaved by one or more selected factors. do.

[0010] In yet another embodiment of the present invention, the cleavable linker is preferably a benzoyl linker. benzyl elimination-based linker ker), trialkyl lock-based linker ed linker, bicine-based linker er), acid labile linker, cleaved in lysosomes Lysosomally cleavable peptide and cathepsin B cleavable peptides and more preferably, Carbamates, disulfides, hydrazines Hydrangea, ester, peptide, Azide, β-Glucuronide and the like The linker is a linker containing any one or more bonds selected from the group consisting of the above combinations. However, it is characterized by its ability to function as a Toll-like receptor 7 or 8 agonist in a cleavable form. They can be chemically bonded to the functional drugs to link them together, and can be activated by endogenous factors (enzymes) in the body. phosphate, redox potential, GSH, pH, etc.) and / or exogenous factors ( redox, pH, temperature, photo / light, magneti c, ultrasound, electrically responsive, etc. There is no limitation thereto so long as the bond can be cleaved.

[0011] In yet another embodiment of the invention, the cleavable linker is at both ends or at one end. One end is ethylene oxide or ethylene glycol. By further incorporating alkyl derivatives such as ethylene glycol , which is characterized by increasing the solubility and flexibility of the conjugate in aqueous solution.

[0012] In yet another embodiment of the invention, the Toll-like receptor 7 or 8 agonist is preferably In particular, imidazoquinoline-based gonist), hydroxyadenine system agonist (8-hydroxyadenine-b ased agonist, pteridone-based agonist, aminopyrimidine- Benzazepine-based agonist based agonists, and thiaoxoguanosine agonists (7-thia 8-oxoguanosine-based agonists The present invention is characterized in that the cleavable linker is attached to the activation site. Toll-like receptor 7 or 8 agonists can exhibit inactivated morphology by If so, this is not limited to this.

[0013] In yet another embodiment of the present invention, the functional drug is preferably an antibody, an antibody fragment, , single chain antibodies, anticancer drugs, antigens, cytokines, proteins, peptides, amino acids, oligonucleotides nucleotides, enzymes, lipids, small molecules, glycoproteins and targeting ligands More preferably, the compound may be any one or more selected from the group consisting of antibodies, proteins, and the like. Any one or more selected from the group consisting of protein-based drugs, sensitizers, and anticancer drugs. Alternatively, a cleavable linker may be used to link the toll-like receptor 7 or 8 agonist. There is no limitation to these, so long as the drug can be combined with the compound.

[0014] The present invention also provides a composition for regulating immune function, which comprises the conjugate as an active ingredient.

[0015] In one embodiment of the present invention, the composition for regulating immune function comprises an antigen-presenting cell, a B cell, a nascent cell, or a cellular component. Any one selected from the group consisting of NK cells and T cells The present invention is characterized by activating one or more immune cells.

[0016] In another embodiment of the present invention, the composition for regulating immune function is tory T cell), MDSC (myeoloid derived suppr essor cells, and M2 macrophages. It is characterized by regulating the function of one or more immune cells.

[0017] In still another embodiment of the present invention, the composition for regulating immune function is administered at an injection site or Activating immune cells in the tumor microenvironment It is possible to induce or control the function of immunosuppressive cells, preferably antigen-presenting cells (d endritic cells, macrophages, etc.), natural killer cells ( Immune cells that induce immune activation such as NK cells and T cells, or have an immunosuppressive effect (Treg(regulatory T cell), MDSC(myeoloid d suppressor cells), M2 macrophage, etc. By regulating the function of , it is possible to regulate immune function in the body.

[0018] In still another embodiment of the present invention, the composition for regulating immune function is an immune checkpoint inhibitor. Anti-PD-1, anti-PD-L1, and anti-CTLA targeting tumor factors -4, anti-KIR, anti-LAG3, anti-CD137, anti-OX 40, anti-CD276, anti-CD27, anti-GITR, anti-T IM3, anti-41BB, anti-CD226, anti-CD40, anti- CD70, anti-ICOS, anti-CD40L, anti-BTLA, anti -TCR, anti-TIGIT, and any one or more selected from the group consisting of It may further comprise an antibody.

[0019] The present invention also relates to a pharmaceutical composition for preventing or treating cancer, comprising the conjugate as an active ingredient. The composition is provided.

[0020] In one embodiment of the present invention, the pharmaceutical composition preferably comprises a chemotherapeutic agent, an immunotherapy agent, or a combination thereof. It may further comprise substances commonly used in the treatment of cancer, such as checkpoint inhibitors. The pharmaceutical composition contains the conjugate of the present invention, thereby effectively activating the immune function in the body. Since it can be activated, it can be used as a therapeutic agent against conventional cancer drugs such as chemotherapy and immune checkpoint inhibitors. It can enhance efficacy.

[0021] In another embodiment of the present invention, the pharmaceutical composition is for use in treating cancer proliferation, metastasis, recurrence or anti-cancer. It is characterized by suppressing resistance to cancer treatment therapies.

[0022] In yet another embodiment of the present invention, the cancer is preferably breast cancer, colon cancer, or rectal cancer. Intestinal cancer, lung cancer, colon cancer, thyroid cancer, oral cancer, pharyngeal cancer, laryngeal cancer, cervical cancer, Brain cancer, ovarian cancer, bladder cancer, kidney cancer, liver cancer, pancreatic cancer, prostate cancer, skin cancer, tongue cancer The cancer may be, but is not limited to, cancer, uterine cancer, stomach cancer, bone cancer, blood cancer, etc. .

[0023] The present invention also includes administering to an individual a composition containing the conjugate as an active ingredient. A method for preventing or treating cancer is provided.

[0024] The present invention also relates to a composition containing the conjugate as an active ingredient for use in the prevention or treatment of cancer. to provide.

[0025] The present invention also relates to a method for producing a drug for preventing or treating cancer comprising the conjugate. Provides uses for.

[0026] The present invention also provides a method for the treatment of toll-like receptor 7 or 8 agonists comprising administering to the toll-like receptor 7 or 8 agonist a cleavable phosphoryl group at the activation site thereof. b) chemically linking a linker to a functional drug; and c) linking the linker to a functional drug. The present invention provides a method for producing a conjugate of a functional drug and a Toll-like receptor 7 or 8 agonist, comprising: Effect of the Invention

[0027] Functional drugs according to the present invention and toll-like receptor 7 or 8 agonists whose activity is temporarily inhibited The conjugates of these drugs are functionalized with Toll-like receptors that can induce synergistic effects with the functional drugs. The second action of the receptor 7 or 8 agonist after a certain period of time enhances nonspecific immunity. Reactions and toxicity can be minimized while synergistic effects can be maximized. In addition, the injection site and tumor microenvironment ), or after delivery into immune cells, functional drugs and toll-like receptor 7 or 8 agonists are gradually separated, and the activation sites of Toll-like receptor 7 or 8 agonists are dynamically modulated ( kinetic immune modulation (IMM) It is possible to minimize the toxicity of 7 or 8 agonists while sustaining a long-term reaction with Toll-like receptors. This leads to a greater inhibition of immune activation of immune cells than the use of Toll-like receptor agonists alone. It can increase the duration and significantly increase the therapeutic effect. In addition, the toll-like receptor 7 or 8 agonist chemically bound to the functional drug according to the present invention can be used. The therapeutic drug (functional drug-Toll-like receptor 7 or 8 agonist conjugate) binds an antibody to the functional drug. When applied, the targeting specificity of antibodies can be used to increase the efficiency of targeting specific cells and tissues. This significantly reduces the side effects and cytotoxicity of conventional Toll-like receptor 7 or 8 agonists. It can also reduce the death of cancer cells by specifically targeting them with antibodies. Cancer antigens produced by IL-1 bind to the adjuvant function of Toll-like receptor 7 or 8 agonists This can also induce the so-called in-situ vaccination effect. Moreover, functional drug-Toll-like receptor 7 or 8 agonist conjugates bind to antigen-presenting cells (APCs). (dendritic cells, macrophages, etc.), natural killer cells It can also induce immune activation of NK cells, T cells, etc., and is effective at the injection site or Treg (regulatory T) cells act as immunosuppressants in the tumor microenvironment. cell), MDSC(myeoloid derived suppressor c Since it can regulate the functions of the M2 macrophages, Not only can it demonstrate anti-cancer effects on its own, but it can also be used as an immune checkpoint inhibitor and chemoattractant. When a conjugate is produced using a functional drug such as a cancer drug, the synergistic effect is effective in preventing the Thus, the conjugates of the present invention can significantly increase the cancer efficacy of toll-like receptors. It can be effectively applied to all functional drugs that can be administered in combination with body 7 or 8 agonists to reduce side effects. It can reduce the amount of stimulants and improve the therapeutic effect, so it can be widely used in various fields. It is expected that... [Brief description of the drawings]

[0028] [Figure 1] FIG. 1 is a diagram showing a representative linker for binding a Toll-like receptor 7 or 8 agonist to a functional drug according to one embodiment of the present invention. [Diagram 2] FIG. 2 is a schematic diagram showing a representative anticancer drug containing an amine group according to one embodiment of the present invention, an example of a conjugate between an anticancer drug containing an amine group having a bond that is cleaved by a reducing agent and a Toll-like receptor 7 or 8 agonist, and the mechanism of cleavage by a reducing agent. [Diagram 3] FIG. 3 is a schematic diagram showing an example of a conjugate between an anticancer drug containing an amine group having a bond that is cleaved under acidic conditions and a Toll-like receptor 7 or 8 agonist according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 4]FIG. 4 is a simplified diagram showing a representative anticancer drug containing a hydroxy group according to one embodiment of the present invention, an example of a conjugate between an anticancer drug having a bond cleaved by β-glucuronidase and a Toll-like receptor 7 or 8 agonist, and the mechanism of cleavage. [Diagram 5] FIG. 5 is a schematic diagram showing an example of a conjugate between an anticancer drug containing a hydroxy group having a bond cleaved by a protease and a Toll-like receptor 7 or 8 agonist according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 6] FIG. 6 is a simplified diagram illustrating representative linkers and linking mechanisms for the synthesis of conjugates between protein-based drugs and Toll-like receptor 7 or 8 agonists according to one embodiment of the present invention. [Figure 7] FIG. 7 is a simplified diagram illustrating an example of a protein-based drug to Toll-like receptor 7 or 8 agonist conjugate having a bond that is cleaved by a reducing agent according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 8] FIG. 8 is a simplified diagram illustrating an example of a conjugate between a protein-based drug and a Toll-like receptor 7 or 8 agonist having a bond that is cleaved by intracellular hydrogen ion concentration in accordance with one embodiment of the present invention, and the mechanism of cleavage. [Figure 9] FIG. 9 is a simplified diagram illustrating an example of a protein-based drug-toll-like receptor 7 or 8 agonist conjugate having a bond that is cleaved by β-glucuronidase according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 10] FIG. 10 is a simplified diagram illustrating an example of a protein-based drug-toll-like receptor 7 or 8 agonist conjugate having a bond that is cleaved by a reducing agent according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 11] FIG. 11 is a simplified diagram illustrating an example of a conjugate between a protein-based drug and a Toll-like receptor 7 or 8 agonist having a bond that is cleaved by intracellular hydrogen ion concentration in accordance with one embodiment of the present invention, and the mechanism of cleavage. [Figure 12]FIG. 12 is a simplified diagram illustrating an example of a protein-based drug-toll-like receptor 7 or 8 agonist conjugate having a bond that is cleaved by β-glucuronidase according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 13] FIG. 13 is a simplified diagram showing representative types of sensitizers according to one embodiment of the present invention, examples of conjugates between sensitizers and Toll-like receptor 7 or 8 agonists having bonds that are cleaved by reducing agents, and the mechanism of cleavage. [Figure 14] FIG. 14 is a schematic diagram showing an example of a conjugate between a sensitizer containing a carboxyl group having a bond that is cleaved under acidic conditions and a Toll-like receptor 7 or 8 agonist according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 15] FIG. 15 is a simplified diagram showing an example of a conjugate between a sensitizer containing a carboxyl group having a bond cleaved by β-glucuronidase and a Toll-like receptor 7 or 8 agonist according to one embodiment of the present invention, and the mechanism of cleavage. [Figure 16] FIG. 16 shows the results of confirming the anti-cancer effect of the AAC-001 conjugate according to one embodiment of the present invention based on the size of the cancer. [Figure 17] FIG. 17 is a diagram showing the results of confirming the anti-cancer effect of the AAC-001 conjugate according to one embodiment of the present invention in terms of survival rate. [Figure 18] FIG. 18 shows the results of confirming the effect of an AAC-001 conjugate according to one embodiment of the present invention on the secretion of inflammatory cytokines in blood. [Figure 19] 19 is a schematic diagram showing the mechanism of the AAC-002 conjugate according to one embodiment of the present invention. More specifically, it is a schematic diagram showing the intracellular mechanism of action of the conjugate of an antibody and a Toll-like receptor 7 or 8 agonist, and shows the process in which the inactive Toll-like receptor 7 or 8 agonist is converted to an active state by the separation of the antibody and the Toll-like receptor 7 or 8 agonist by the intracellular environment. [Figure 20] FIG. 20 shows the results of confirming the effect of an AAC-002 conjugate according to one embodiment of the present invention on the secretion of inflammatory cytokines in blood. [Figure 21] FIG. 21 shows the results of confirming that a conjugate of resiquimod and a protein antigen having a bond that can be cleaved by cathepsin B forms nanoparticles of about 99.7 nm in an aqueous solution. [Figure 22] Figure 22 is a schematic diagram showing the structure and function of a conjugate of a functional drug and a Toll-like receptor 7 or 8 agonist, and shows the difference between kinetically activating conjugates, in which a functional drug is bound to the activation site of a Toll-like receptor 7 or 8 agonist by a cleavable linker and temporarily inactivated, and conventional conjugates, in which a functional substance is bound to another site rather than the activation site. Kinetically activating conjugates are characterized in that only the efficacy of the functional substance is effective initially, and after the cleavable linker reacts, the efficacy of the inactivated Toll-like receptor 7 or 8 agonist is secondarily kinetically regenerated in response to a stimulus, whereas conventional conjugates, which maintain the dual active state of the functional substance function and the Toll-like receptor agonist from the beginning, cannot embody such a kinetic action. BEST MODE FOR CARRYING OUT THEINVENTION

[0029] The present inventors demonstrated that Toll-like receptor 7 or 8 agonists suppress non-specific early immune responses. This reduces side effects and allows various functional drugs to be combined for one-time administration. To prepare a conjugate of a functional drug and a Toll-like receptor 7 or 8 agonist that can be administered in combination. As a result of intensive research, we have developed a method to insert a cleavable linker into the activation site of Toll-like receptor 7 or 8 agonists. By binding a functional drug using the The toll-like receptor agonist, whose immune activation activity is temporarily inhibited, is injected into the tumor microenvironment. Recovery of immune activation function specifically in the environment or target cells Secondary acting conjugates were developed (Figure 22). The inhibition was observed in Toll-like receptors 7 or 8. It may also mean that the function of the agonist's active site is delayed. The conjugate of the present invention between the functional drug and the toll-like receptor 7 or 8 agonist can be used in the conventional binding. Compared with conventional conjugates, functional drugs and toxins kinetically modulate the activity of agonists of leukocyte-like receptors 7 and 8 The difference is that it can be dynamically modulated. be.

[0030] As used herein, the term "toll-like receptor agonist" refers to a toll-like receptor agonist. "Tor-like receptor agonist" is a membrane protein involved in innate immunity. Direct or indirect acting ligands, which are signaling pathways mediated by endogenous or exogenous ligand production. It may refer to a component capable of initiating a signal transduction reaction via a neutral transduction pathway. In the present specification, the toll-like receptor agonist refers to a natural toll-like receptor agonist or a synthetic toll-like receptor agonist. The toll-like receptor agonist may be a toll-like receptor 1 agonist, a toll-like receptor 2 agonist, a toll-like receptor 3 agonist, a toll-like receptor 4 agonist, a toll-like receptor 5 agonist, a toll-like receptor 6 agonist, a toll-like receptor 7 agonist, a toll-like receptor 8 agonist, a toll-like receptor 9 agonist, a toll-like receptor 1 Toll-like receptor 3 agonist, Toll-like receptor 4 agonist, Toll-like receptor 5 agonist, Toll-like receptor The toll-like receptor 7 or 8 agonist, the toll-like receptor 9 agonist, etc. TLR-7 or -8 agonists are ligands that can induce signaling responses via TLR-7 or -8. An example of such an agent is an imidazoquinolone. ne-based agonist, hydroxyadenine agonist (8-hydrox Yadenine-based agonist, pteridone agonist one-based agonist, aminopyrimidine agonist (2-aminop yrimidine-based agonist, benzazepine agonist zoazepine-based agonist), thiaoxoguanosine agonist ( 7-thia-8-oxoguanosine-based agonist) The imidazoquinoline compound may be any of those described in WO2018 196823 and WO20 11 049677, WO2011 027022, WO2017 102652, WO 2019 040491, etc., or a pharma- ceutical acceptable salt thereof. However, the present invention is not limited thereto. In addition, the hydroxyadenine compound may be 80730, WO2013 068438, WO2019 036023, WO2019 035969, WO2019 035970, WO2019 035971, WO20 19 035968, CN 108948016, US 2014 8846697, W O2016 023511, WO2017 133683, WO2017 133686 , WO2017 133684, WO2017 133687, WO2017 0763 46, WO2018 210298, WO2018 095426, WO2018 06 8593, WO2018 078149, WO2018 041763, etc. The pteridone class includes, but is not limited to, compounds of the above-mentioned type or pharma- ceutically acceptable salts. The compounds are disclosed in US 2010 0143301, WO2016 007765, WO20 16 044182, WO2017 035230, WO2017 219931, WO Compounds of the same type as those described in CN 2011 057148, CN 1087 94486, etc. or a pharma- ceutically acceptable salt thereof. WO2010 133885, WO2012066335, WO2012 0663 36, WO2012 067268, WO2013 172479, WO2012 13 6834, WO2014 053516, WO2014 053595, US 2018 0215720, WO2012 156498, WO2014 076221, WO2 016 141092, WO2018 045144, WO2015 014815, W O2018 233648, WO2014 207082, WO2014 056593 , WO2018 002319, WO2013 117615, etc. The benzazepine compounds include, but are not limited to, benzoazepine derivatives, their derivatives, and pharma- ceutically acceptable salts. The compound is disclosed in WO2007 024612, WO2010 014913, and WO2010 0 54215, WO2011 022508, WO2011 022509, WO2012 097177, WO2012 097173, WO2016 096778, WO20 16 142250, WO2017 202704, WO2017 202703, WO 2017 216054, WO2017 046112, WO2017 197624, etc. The present invention includes, but is not limited to, compounds of the type described above or pharma- ceutically acceptable salts thereof. The thiaoxoguanosine-based compound is disclosed in WO2016 180691, WO2016 0 55553, WO2016 180743, WO2016 091698, etc. The present invention includes, but is not limited to, compounds of the above-mentioned type or pharma- ceutically acceptable salts. PCT / US2009 / 035563, PCT / US2015 / 028264, PCT / US2016 / 020499, WO2015 023598, PCT / US 2015 / Toll-like receptor 7 or 8 compounds or pharma- ceutical acceptable salts thereof, as described in 039776, etc. It may contain salts. Also, Imiquimod, Resiquimod uimod, Dactolisib, Gardiq uimod, Sumanirole, Motolimo d), vesatolimod, loxoribine ), SM360320, CL264, 3M-003, IMDQ, Compound 54 However, the present invention is not limited to these examples, and may be used by those skilled in the art. This includes all cases of toll-like receptor 7 or 8 agonists that can inhibit the toll-like receptor 7 or 8. The agonist is a toll-like receptor agonist. nist), saponin, antiviral peptide, inflammasome inducer (inf lammasome inducer), NOD ligand, C DS ligand (cytosolic DNA sensor ligand), STIN G (stimulator of interferon genes) ligand, Emma Emulsion, alum and combinations thereof It can be replaced with one or more immune activators selected from the group, and delivered into cells to activate the Toll-like receptor 3 agonists or Toll-like receptor 9 agonists that have receptors inside the endosome The same concept can be applied to drugs to synthesize antibodies and conjugates, but is not limited thereto. stomach.

[0031] As used herein, the term "cleavable linker" refers to a It contains a cleavable bond and is expressed in the tumor microenvironment, intracellular endosomes and lysosomes. the physiological environment of the body, such as low pH, enzymes, glutathione, etc.; or external Stimuli, namely temperature, redox potential, ultrasound, Specific stimuli such as magnetic fields and near-infrared light The term "linker" generally refers to a linker that can be cleaved by te), disulfide, hydrazine, ethene Ester, peptide, azide, β-glucose It means a linker containing a bond such as β-Glucuronide. The conjugate may be in any form as long as it is cleavable. Various enzymes, acid phosphatase, and phosphatase inhibitors present at the site of tumor injection, in the tumor microenvironment, or within cells ase, Acid phyrophosphatase, Phosphodiester ase, Phosphoprotein phosphatase, Phosphati dic acid phosphatase, Arylsulfatase, Prote ases, Cathepsins, Collagenase, Arylamidase, Peptidase, Acid ribonuclease, Acid deoxyri bonuclease, Lipases, Triglyceride lipase, P Hospholipase, Esterase, Carboxyesterase, Cl ucocerebrosidase, Galactocerebrosidase, Sp hingomyelinase, Glycosidases, alpha-Glucos idase, beta-Glucosidase, beta-Galactosidas e, alpha-Mannosidase, alpha-ucosidase, beta -Xylosidase, alpha-N-Acetylhexosaminidase , beta-N-Acetylhexosaminidase, Sialidase, L ysozyme, Hyaluronidase, beta-Glucuronidase , gamma-interferon-inducible lysosomal th iol reductase and other drugs that act as functional drugs and toll-like receptor 7 or 8 agonists It can also be separated.

[0032] As used herein, a "functional drug" refers to a drug that is effective in treating a disease. Anything that has a biological function that can be used for diagnosis, prevention, treatment, etc. The above drugs are generally referred to as antibodies, antibody fragments, single-chain antibodies, anticancer drugs, antigens, cytokines, etc. Insulin, proteins, peptides, amino acids, oligonucleotides, enzymes, lipids, small molecule compounds The antibody may be a peptide, a glycoprotein, a targeting ligand, or the like, and more preferably an antibody. The therapeutic agent may be, but is not limited to, a cancer drug, an antibody, a protein-based drug, a sensitizer, etc. Examples of the functional drug include DNA topoisomerase inhibitors (DNA topoisomerase inhibitor, microtubule inhibitor ule inhibiting drug, DNA damaging agent agent), antimetabolite, nucleoside analogue (nucleoside analog), antisense oligonucleotide, lock nucleus Acid (locked nucleic acid; LNA), small interfering RNA (siRN) A; short interfering RNA, microRNA (microRNP) A; miRNA, aptamer, peptide nucleic acid nucleic acid (PNA), phosphorodiamidate morpholino oligonucleotide phosphorodiamidate morpholino oligonuc leotides;PMO), antisense Bcl-2 oligonucleotides, antisense Antisense HIF-1α oligonucleotides, antisense Survivin oligonucleotides nucleotides, cell adhesion peptides, Cell penetrating peptide, receptor linker receptor ligand, targeting carbohydrate molecule carbohydrate molecule, lectin, RGD Peptide, selectin, TAT, penetratin In, (Arg)9, folic acid, etc. Microtubulin synthesis inhibitors used in treatment , meiosis inhibitors, RNA polymerase inhibitors, topoisomerase poisomerase inhibitors, DNA intercalators ators, DNA alkylators, ribosome inhibitors , protein toxins, radioisotopes, etc. Also, maytansinoids (maytansinoids) ansinoid, auristatin, dolastatin lastatin, tubulysin, calicheamicin Icheamicin, pyrrolobenzodiazepine epines), doxorubicin, duocarmycin uocarmycin), carboplatin (paraplatin) paraplatin), cisplatin, cyclophosphamide (cyclophosphamide), ifosfamide, Nidran, nitrogen mustard (mechlorethamine hydrochloride) rogen mustar (mechlorethamine HCL)], bleomycin Bleomycin, mitomycin C, cytarabi Cytarabine, fluorouracil, gemcitabine Gemcitabine, trimetrexate , methotrexate, etoposide , vinblastine, vinorelbine ), alimta, altretamine, procal Procarbazine, Taxol, Taxotere xotere, topotecan, irinotecan an), trichothecene, CC1965, alpha-ama alpha-amanitin, other enediyne antibiotics, plant toxins, etc. In the case of a compound, the form of its stereoisomer or derivative may also be included. The auristatin is monomethyl auristatin E (monomethyl auristatin E). statin E) or monomethyl auristatin F istatin F), but any substance used in cancer treatment There are no restrictions. In addition, protoporphrin, hematoporp hrin monomethryl ether, pheophorbide A, ph otofrin, etc., can significantly accelerate chemical reactions and physical phenomena. However, the present invention is not limited to these, so long as the chemical sensitizer or optical sensitizer can be used.

[0033] As used herein, the term "antibody" refers to a substance that is immunologically reactive with a specific antigen. It acts as a receptor for antigens that specifically recognize antigens, including immunoglobulin molecules having the following structure: It refers to a protein molecule, and can be used for polyclonal antibodies, monoclonal antibodies, full-length antibodies, etc. th) Includes whole antibodies and antibody fragments that contain the antigen-binding domain. Full-length antibodies are composed of two whole The structure has a full-length light chain and two full-length heavy chains, each of which is The whole antibodies are linked by disulfide bonds. and IgG, which is a subtype, including IgG1, IgG2, The antibody fragment includes IgG3 and IgG4. The antibody fragment means a fragment that retains the antigen-binding function. The Fab fragments include Fab, Fab', F(ab')2, scFv and Fv. , the light chain and heavy chain variable regions, the light chain constant region, and the first constant region of the heavy chain (CH1) Fab' is a structure that contains one antigen-binding site. has a hinge region containing one or more cysteine ​​residues. F(ab')2 antibodies differ from Fab in that they have a structure similar to that of the hinge region of Fab'. Fv (variable fragment length) is formed by disulfide bonds between the amino acid residues. ent) refers to the smallest antibody fragment that contains only the heavy chain variable region and the light chain variable region. In double-chain Fv (dsFv), the heavy chain variable region and the light chain variable region are linked by a disulfide bond. Single-chain Fv (scFv) is generally composed of a heavy chain variable region linked via a peptide linker. The antibody fragments are composed of a covalent bond between the variable domain and the light chain variable domain of the protein. It can be obtained using hydrolases (e.g., whole antibodies can be subjected to limited digestion with papain to obtain Fa b can be obtained, and when digested with pepsin, F(ab')2 fragments can be obtained. Preferably, the antibody of the present invention can be produced by recombinant gene technology. It can be a natural antibody or a recombinant antibody. A natural antibody means an antibody that has not been genetically engineered. In addition, the immunogenicity that genetically engineered antibodies can have in vivo The risk of viral infection is significantly lower. Recombinant antibodies refer to genetically engineered antibodies. They have the advantage that they can be genetically engineered to add antigen-binding properties or other desired characteristics. Examples of antibodies include , DEC205, CD206, DC-SIGN, DNGR1, CD11c, FcγR, P D-L1, PD-1, CD47, SIRPalpha, Ly6G, IL-6, Gr-1 etc. The antibody may be an antibody that specifically binds to an immune checkpoint inhibitor. checkpoint inhibitors) anti-PD-1, anti-P D-L1, anti-CTLA-4, anti-KIR, anti-LAG3, anti -CD137, anti-OX40, anti-CD276, anti-CD27, an ti-GITR, anti-TIM3, anti-41BB, anti-CD226, a nti-CD40, anti-CD70, anti-ICOS, anti-CD40L, It may be anti-BTLA, anti-TCR, anti-TIGIT, etc.

[0034] In the present specification, the term "drug for controlling immune function" refers to a combination of the functional drug of the present invention and Toll-like receptor 7. or a composition comprising a conjugate of an agonist as an active ingredient, which activates immune cells and It is a drug that can regulate the function of immune suppressor cells and allow the body's immune system to function normally. do.

[0035] As used herein, "prevention" refers to the administration of a composition according to the present invention. It refers to any action that suppresses or delays the onset of diseases such as cancer through the administration of medicine.

[0036] As used herein, "treatment" refers to the administration of a composition according to the present invention. It refers to any action that improves or beneficially alters symptoms such as cancer.

[0037] As used herein, "individual" or "subject" refers to a This refers to a subject to which the composition of the invention can be administered, and there is no limitation on the subject.

[0038] As used herein, "cancer" refers to a cancer that develops locally through invasion and metastasis. It is a general term for various blood cancers, malignant solid tumors, etc. that can be expanded more systematically. Specific examples of cancer include colon cancer, adrenal cancer, bone cancer, brain cancer, breast cancer, Bronchial cancer, colon and / or rectal cancer, gallbladder cancer, gastrointestinal cancer, head and neck cancer, Kidney cancer, laryngeal cancer, liver cancer, lung cancer, nervous tissue cancer, pancreatic cancer, prostate cancer, parathyroid cancer Other examples of cancer include adenocarcinoma, Adenoma, basal cell carcinoma, cervical dysplasia and intraepithelial carcinoma, Ewing sarcoma , squamous cell carcinoma, esophageal cell carcinoma, malignant brain tumor, esophageal cell carcinoma, intestinal ganglioneuroma, hyperplasia Corneal nerve cancer, islet cell carcinoma, Kaposi's sarcoma, leiomyoma, leukemia, lymphoma , malignant carcinoma, malignant melanoma, malignant hypercalcemia, Marfan habitus id habitus cancer, medullary carcinoma, metastatic skin cancer, mucosal neuroma, myelodysplastic syndrome group, myeloma, mycosis fungoides, neuroblastoma, osteosarcoma, osteogenic and other sarcomas, ovarian cancer, Pheochromocytoma, polycythemia vera, primary brain tumors, small cell lung cancer, ulcerative and Papillary squamous cell carcinoma, seminoma, soft tissue sarcoma, retinoblastoma, renal cell tumor or renal cell carcinoma ( renal cell carcinoma (RCC), reticulum cell sarcoma, and Wilms' disease Also included are astrocytomas and gastrointestinal stromal tumors. al stromal tumor, GIST), glioma or glioblastoma, hepatocellular carcinoma Hepatocellular carcinoma (HCC), pancreatic endocrine cancer, etc. etc. are included.

[0039] As used herein, the term "pharmaceutical composition" refers to a "Medicine (production)" refers to the form of capsules, tablets, granules, injections, ointments, powders or beverages. The pharmaceutical composition is characterized in that it is intended for humans. The target composition can be prepared by a conventional method, such as, but not limited to, powder, granules, etc. Granules, capsules, tablets, oral dosage forms such as aqueous suspensions, topical preparations, suppositories and sterile injection solutions The pharmaceutical composition of the present invention may be formulated and used in the form of a pharma- ceutical acceptable The pharmaceutical composition may contain a carrier. Pharmaceutically acceptable carriers include binders, lubricants, and Disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, colorants, flavors, etc. may be used. In the case of injections, it is possible to use buffering agents, preservatives, analgesics, solubilizers, isotonicity agents, stabilizing agents, etc. For topical administration, the drug may be used in combination with a base, excipient, lubricant, etc. Preservatives and the like can be used. The dosage form of the pharmaceutical composition of the present invention can be prepared by adding the above-mentioned pharmaceutical agent. It can be prepared in various forms by mixing with a pharma- ceutical acceptable carrier. For example, It is available in tablets, troches, capsules, elixirs, suspensions, and siro. In the case of injections, they can be manufactured in the form of unit dose ampoules. Alternatively, it may be prepared in a multiple dose form. It can be formulated as a sustained release preparation.

[0040] On the other hand, examples of carriers, excipients and diluents suitable for formulation include lactose, dextrose, and the like. Lose, sucrose, sorbitol, mannitol, xylitol, erythritol, malate Lutitol, starch, gum acacia, alginate, gelatin, calcium phosphate , calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl chloride Nylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc , magnesium stearate or mineral oil may also be used. They may further contain a flocculant, a lubricant, a moisturizer, a flavoring, an emulsifier, a preservative, and the like.

[0041] The administration route of the pharmaceutical composition according to the present invention includes, but is not limited to, oral, Intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, percutaneous, subcutaneous, intraperitoneal, intranasal, intestinal The term "oral" refers to a pharmaceutical composition that is administered orally, topically, sublingually or rectally. Oral or parenteral administration is preferred. The term "parenteral" includes any of the following: subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intrasynovial, intrathoracic, intravenous, intravenous, intravenous. This includes intraosseous, intradural, intralesional and intracranial injection or infusion techniques. It may also be administered in the form of suppositories for rectal administration.

[0042] The pharmaceutical compositions of the present invention may be prepared in a variety of ways, depending on the activity of the particular compound used, age, weight, general health, Gender, diet, time of administration, route of administration, excretion rate, drug combination and specific disease being prevented or treated The dosage of the pharmaceutical composition may vary depending on various factors, including the severity of the disease. The dosage varies depending on the patient's condition, weight, degree of illness, drug form, route of administration and duration. The supplier can select the appropriate dose, 0.0001 to 500 mg / kg or 0.0 The dose can be 0.1 to 500 mg / kg. It can also be administered once a day. The dosages mentioned above are within the scope of the present invention in all respects. The pharmaceutical composition according to the present invention may be in the form of a pill, a sugar-coated tablet, a capsule, a liquid, It can be formulated as a gel, syrup, slurry, or suspension.

[0043] In the following, preferred embodiments are presented to aid in understanding the present invention. The following examples are provided so that the present invention may be more readily understood. Therefore, the contents of the present invention are not limited thereto.

[0044] [Example] Example 1: 2-Methyl-1-(3-nitroquinolin-4-ylamino)propane-2- Synthesis of ol 2-Methyl-1-(3-nitroquinolin-4-ylamino) ) propan-2-ol (compound 2) was synthesized. More specifically, compound 2 was synthesized at 10 to 20°C. In dichloromethane (450 ml) containing 1 (30 g), 1-amino-2-methylpropane Pan-2-ol (14 g) and tetraethylamine (9.6 g) were added and stirred for 2 hours. The mixture was then concentrated by evaporating the solvent under vacuum. Afterwards, it was resuspended in methyl tert-butyl ether (150 ml). The mixture was separated using a filter and then concentrated under low pressure to give compound 2 (32 g, The obtained compound 2 has the following structure: 1 Using H NMR It has been verified. 1 H NMR (400MHz, DMSO-d6): δ9.91 (brs, 1H), 9. 18(s, 1H)), 8.46(d, J=8.0Hz, 1H), 7.83-7.92(m , 2H), 7.56-7.60(m, 1H), 5.15(s, 1H), 3.86(d, J =4.8Hz, 2H), 1.15(s, 6H).

[0045] [ka]

[0046] Example 2: 1-(3-aminoquinolin-4-ylamino)-2-methylpropane-2- Synthesis of ol Using the method of the following reaction scheme 2, 1-(3-aminoquinolin-4-ylamino)-2-methyl More specifically, dipropan-2-ol (compound 3) was synthesized at a reaction temperature of 10 to 20°C. Compound 2 (32 g), methanol (500 ml), and Pd / C catalyst (3.2 g After mixing, the mixture was degassed and flushed three times with hydrogen. The hydrogen was vaporized and the pressure was kept at 1 atm. After that, the mixture was stirred at room temperature for 5 hours. Then, the mixture was diluted with methyl tert-butyl ether (1 The resuspended mixture was separated using a filter, Concentration under low pressure gave compound 3 (27 g, 95.4%, yellow solid). The structure of compound 3 is: 1 This was verified using H NMR. 1 H NMR (400MHz, DMSO-d6): δ8.37(s, 1H)), 7.9 9-8.01(m, 1H), 7.72-7.74(m, 1H), 7.32-7.39(m , 2H), 5.04(s, 2H), 4.77(brs, 1H), 4.67-4.70(m , 1H), 4.12(brs, 2H), 1.15(s, 6H).

[0047] [ka]

[0048] Example 3: 1-(2-ethoxymethyl)-1H-imidazo[4,5-c]quinoline-1 Synthesis of (-yl)-2-methylpropan-2-ol Using the method shown in Reaction Scheme 3 below, 1-(2-ethoxymethyl)-1H-imidazo[4,5- c]Quinolin-1-yl)-2-methylpropan-2-ol (Compound 4) was synthesized. More specifically, compound 3 (27 g) and 2-ethoxyacetic acid were added to a reactor at 10 to 20° C. (30 ml) was added, and the mixture was stirred at 120 to 130°C for 5 hours. The mixture was cooled to 20-25°C and saturated sodium carbonate (150 ml) was added. The reaction mixture was extracted with a mixture of chloromethane and methanol (10 / 1, v / v). The extracted organic layer was washed with brine and then diluted with sodium sulfate (10 g). The organic solution layer from which the water had been removed was then filtered using a filter. Compound 4 (30 g, 85.8%, yellow gel) was obtained by filtering and concentrating under low pressure. The structure of the obtained compound 4 is: 1 This was verified using H NMR. 1 H NMR (DMSO_d6 400MHz): δ9.18(s, 1H)), 8.63 (d, J=8.0Hz, 1H), 8.13(dd, J=1.6, 8.0Hz, 1H), 7 .63-7.71(m, 2H), 4.91(s, 2H), 4.78(brs, 2H), 3 .54(q, J=6.8Hz, 2H), 1.10-1.18(m, 9H).

[0049] [ka]

[0050] Example 4: 2-(ethoxymethyl)-1-(2-hydroxy-2-methylpropyl)- Synthesis of 1H-imidazo[4,5-c]quinoline 5-oxide Using the method of the following reaction scheme 4, 2-(ethoxymethyl)-1-(2-hydroxy-2-methyl) (ethylpropyl)-1H-imidazo[4,5-c]quinoline 5-oxide (compound 5) More specifically, compound 4 (30 g), dichloromethane, and 1,2-dichloromethane were added to a reactor at 10 to 20°C. After adding 20 ml of benzoic acid (350 ml) and m-chloroperoxybenzoic acid (26 g), the mixture was cooled to room temperature. The mixture was stirred at room temperature for 4 hours. Then, saturated sodium carbonate solution (150 ml) and Sodium sulfate solution (150 ml) was added, followed by dichloromethane and methanol (10 / 10). The reaction product was extracted using a mixed solution of 1, v / v). The water in the extracted organic solution layer was removed with sulfuric acid. After removing sodium (30 g), the mixture was filtered. The reaction mixture was concentrated under low pressure. The concentrated reaction mixture was then diluted with ethyl acetate (50 ml). The mixture was resuspended in water, separated using a filter, and dried under low pressure to obtain compound 5 (30 The obtained compound 5 has the following structure: 1 Using H NMR We verified this by: 1 H NMR (DMSO_d6 400MHz): δ9.04(s, 1H)), 8.79 (d, J=8.4Hz, 1H), 8.71(d, J=8.4Hz, 1H), 7.77-7 .80(m, 2H), 4.93(s, 2H), 4.73(brs, 2H), 3.54(q , J=6.8Hz, 2H), 1.12-1.18(m, 9H).

[0051] [ka]

[0052] Example 5: 1-(4-amino-2-ethoxymethyl)-1H-imidazo[4.5-c] Synthesis of quinolin-1-yl)-2-methylpropan-2-ol 1-(4-amino-2-ethoxymethyl)-1H-imidazolium chloride was prepared by the method shown in Reaction Scheme 5 below. Zo[4.5-c]quinolin-1-yl)-2-methylpropan-2-ol (compound 6) More specifically, compound 5 (30 g), DCM ( 600 ml), 4-methylbenzene-1-sulfonyl chloride (18.2 g), and Aqueous ammonia (NH3·H2O, 180 ml) was added and the mixture was stirred at room temperature for 16 hours. Next, distilled water was added to the stirred mixture, followed by dichloromethane and methanol (10 / 1, The mixture was separated using a v / v mixture of 100% ethanol and 100% ethanol. The separated organic layer was washed with brine. After washing with e), the water was removed using anhydrous sodium sulfate (50 g). The organic solution layer from which the benzene has been removed is filtered and then concentrated under low pressure. The reaction mixture was concentrated and diluted with methyl tert-butyl ether and methanol (15 / 1, v The cells were then resuspended in a mixed solution of 1000 mM NaCl (1:100 / v) for 30 minutes. After that, the cells were separated using a filter. The mixture was dried under low pressure to obtain compound 6 (18 g, 60%, yellow solid). The structure of compound 6 is: 1 This was verified using H NMR. 1 H NMR (DMSO_d6 400MHz): δ8.27(d, J=8.0Hz, 1 H), 7.59(d, J=7.6Hz, 1H), 7.40(t, J=7.2Hz, 1H) , 7.21(t, J=7.2Hz, 1H), 6.57(brs, 2H), 4.89(s, 2H), 4.68(brs, 2H), 3.52(q, J=6.8Hz, 2H), 1.11 -1.17(m, 9H).

[0053] [ka]

[0054] Example 6: 10,13-Dimethyl-17-(6-methylheptan-2-yl)-2,3 ,4,7,8,9,10,11,12,13,14,15,16,17-Tetradecahydride 1H-cyclo[a]phenanthren-3-yl 2-(ethoxymethyl)-1-(2- Hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-yl Carbamate synthesis 10,13-dimethyl-17-(6-methylheptane-2) -Il)-2,3,4,7,8,9,10,11,12,13,14,15,16,17 -Tetradecahydro-1H-cyclo[a]phenanthren-3-yl 2-(ethoxymethyl) (2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]ky We synthesized 4-norin-4-yl carbamate (compound 8). First, compound 7 (TCI, 50 g, Cholesterol chloroformate) in 250 g of silica gel Column chromatography packed with 0% to 20% ethyl acetate in n-hexane Using this method, pure compound 7 (30 g) was obtained. Compound 7 was then added to a reactor at 10-20°C. After addition of 6 (15 g) and dichloromethane (198.9 g), the pure compound 7 (30 g) was ) and tetraethylamine (9.6 g) were added in that order, and the mixture was stirred at 20 to 25° C. for 16 hours. Water was added to the stirred mixture, followed by the addition of dichloromethane to extract the reaction. The organic layer was washed with salt water and then extracted with anhydrous sodium sulfate (195 g). The water was removed. The organic solution layer from which the water had been removed was filtered using a filter. The reaction mixture was then concentrated under reduced pressure. The cells were then resuspended in a mixture of ether and methanol (10 / 1, v / v). After separation using a cyclohexane separator, the mixture was dried under low pressure to obtain compound 8 (10.2 g, 55.1%, The obtained compound 8 has the following structure: 1 This was verified using H NMR. 1 Through the H NMR results, resiquimod (R848) and cholesterol It was confirmed that a conjugate in which the amine chain was linked to the amine chain via a carbamate bond was prepared. 1 H NMR (CDCl3400MHz): δ8.13-8.19(m, 2H), 7 .59-7.63(m, 1H)), 7.46-7.50(m, 1H), 5.42-5.4 3(m, 1H), 4.92(brs, 2H), 4.72-4.80(m, 3H), 3.6 8(q, J=6.8Hz, 2H), 3.24(s, 1H), 2.51-2.59(m, 1 H), 2.36-2.47(m, 1H), 1.96-2.11(m, 3H), 1.81- 1.95(m, 2H), 1.45-1.75(m, 9H), 1.02-1.35(m, 2 7H), 0.94(d, J=6.4Hz, 3H), 0.89(d, J=6.4Hz, 6H ), 0.71(s, 3H).

[0055] [ka]

[0056] Example 7: Bis(2,5-dioxopyrrolidin-1-yl) 2,2'-disulfane di Synthesis of isopropylbis(ethane-2,1-diyl)dicarbonate Bis(2,5-dioxopyrrolidin-1-yl) 2,2' is synthesized by the method shown in Reaction Scheme 7 below. - Disulfanediylbis(ethane-2,1-diyl)dicarbonate (compound 9) was synthesized. First, compound 7 (TCI, 70 g) was added to a column packed with 350 g of silica gel. The pure compound was obtained by chromatography (0% to 20% ethyl acetate in n-hexane). Compound 7 (40 g) was obtained. Then, pure compound 7 (40 g) was added to the reactor at 10-15 °C. ) and dichloromethane (100 ml), and then bis(2-hydroxyethyl)dichloromethane (100 ml) was added. A solution of sulfide in dichloromethane (250 ml) and pyridine (21 g) was added. The mixture was stirred at room temperature for 2 hours, and then distilled water (200 ml) was added. The reaction mixture was then extracted with three portions of dichloromethane (150 ml). The solution layer was washed with salt water and then dehydrated with anhydrous sodium sulfate (20 g). The reaction mixture from which moisture had been removed was filtered and then placed under low pressure. The mixture was then concentrated by column chromatography (silica gel, 300 g, 10% to 30% Compound 9 (20 g, 39.6%, yellow gel) was extracted with 0% ethyl acetate in n-hexane. The structure of the obtained compound 9 is: 1 This was verified using H NMR. 1 H NMR (CDCl3400MHz): δ5.41-5.42 (m, 1H), 4. 46-4.56(m, 1H), 4.41(t, J=6.8Hz, 2H), 3.91(t, J=6.0Hz, 2H), 2.98(t, J=6.8Hz, 2H), 2.91(t, J= 6.0Hz, 2H), 2.35-2.47(m, 2H), 1.79-2.08(m, 6H) ), 1.43-1.73(m, 7H), 1.25-1.42(m, 5H), 1.06-1 .22(m, 7H), 0.97-1.03(m, 5H), 0.93(d, J=6.4Hz , 3H), 0.88(dd,J=1.6,6.8Hz, 6H), 0.69(s, 3H).

[0057] [ka]

[0058] Example 8: Synthesis of Compound 10 Compound 10 was synthesized by the method of the following reaction formula 8. More specifically, Compound 9 (20 g) and dichloromethane (200 ml) were added to the actor, and then the bis(2 ,5-dioxopyrrolidin-1-yl) carbonate (18g) and tetraethylamine ( The mixture was stirred at room temperature for 3 hours, then distilled water (300 ml) was added. l) was added, and the reaction was then extracted with three additions of dichloromethane (150 ml). The extracted organic solution layer was washed with salt water and then extracted with anhydrous sodium sulfate (20 g). The reaction mixture from which moisture had been removed was filtered using a filter. The filtrate was then concentrated under low pressure. , 200 g, 5% to 20% ethyl acetate in n-hexane) to prepare compound 10 (18 g The obtained structure of compound 10 is: 1 Using H NMR It has been verified. 1 H NMR (CDCl3400MHz): δ5.39-5.40 (m, 1H), 4. 57(t, J=6.8Hz, 2H), 4.43-4.52(m, 1H), 4.37(t, J=6.4Hz, 2H), 2.96-3.03(m, 4H), 2.84(s, 4H), 2 .34-2.44(m, 2H), 1.78-2.04(m, 5H), 1.42-1.73 (m, 7H), 1.22-1.40(m, 5H), 1.06-1.20(m, 7H), 0 .95-1.04(m, 5H), 0.91(d, J=6.0Hz, 3H), 0.86(d , J=6.4Hz, 6H), 0.67(s, 3H).

[0059] [ka]

[0060] Example 9: 2-((2-((10,13-dimethyl-17-(6-methylheptane-2 -Il)-2,3,4,7,8,9,10,11,12,13,14,15,16,17 -Tetradecahydro-1H-cyclo[a]phenanthren-3-yloxy)carbonyl 2-(ethoxymethyl)-1-(2-hydroxyethyl)disulfanyl)ethyl (2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-ylcarbamazepine Synthesis of tetracycline Using the method of the following reaction scheme 9, 2-((2-((10,13-dimethyl-17-(6-methyl- ethylheptan-2-yl)-2,3,4,7,8,9,10,11,12,13,14, 15,16,17-Tetradecahydro-1H-cyclo[a]phenanthrene-3-yl 2-(ethoxymethyl)-1-oxycarbonyloxyethyl disulfanylethyl -(2-hydroxy-2-methylpropyl)-1H-imidazo[4,5-c]quinoline- 4-ylcarbamate (compound 11) was synthesized. More specifically, the reaction was carried out at 10 to 20°C. Compound 6 (15 g) and dichloromethane (198.9 g) were added to the flask, and then compound 10 was added. (40.5 g) and tetraethylamine (9.6 g) were added in that order. After stirring at 5°C for 16 hours, distilled water (225 ml) was added. The reaction mixture was extracted by adding 5 portions of ethanol (99.45 g). The extracted organic layer was washed with brine. After washing with water, the water was removed using anhydrous sodium sulfate (195 g). The reactant from which the ions have been removed is filtered through a filter, and the filtrate is then heated under low pressure. The mixture was concentrated, and then subjected to column chromatography (silica gel, 100 g, 10% to 50% % ethyl acetate in n-hexane) to obtain compound 11 (10.8 g, 37.4%, white The obtained compound 11 has the following structure: 1 This was verified using H NMR. 1 1 H NMR results showed that R848 and cholesterol were cross-linked by disulfide. It was confirmed that the compound was produced. 1 H NMR (CDCl3400MHz): δ8.15-8.17(m, 2H), 7 .60-7.64(m, 1H)), 7.47-7.51(m, 1H), 5.39-5.4 0(m, 1H), 4.93(s, 2H), 4.81(s, 2H), 4.56(t, J=6 .4Hz, 2H), 4.45-4.54(m, 1H), 4.41(t, J=6.4Hz, 2H), 3.68(q, J=6.8Hz, 2H), 3.13(s, 1H), 3.09(t , J=6.4Hz, 2H), 3.01(t, J=6.4Hz, 2H), 2.34-2.4 7(m, 2H), 1.92-2.06(m, 3H), 1.79-1.90(m, 2H), 1.23-1.72(m, 21H), 1.06-1.21(m, 7H), 0.96-1. 05(m, 5H), 0.93(d, J=6.4Hz, 3H), 0.88(dd,J=1. 6,6.4Hz, 6H), 0.69(s, 3H).

[0061] [ka]

[0062] Example 10: Synthesis of Toll-like receptor 7 or 8 agonist-antibody conjugates Diverse Toll-like receptors conjugated with antibodies Antibodies that act on the 7th and 8th amino acids (imidazoquinoline-based antibodies) based agonist, hydroxyadenine based agonist (8-hydroxyad enine-based agonist, pteridone -based agonist, aminopyrimidine agonist (2-aminopyri midine-based agonist, benzazepine zepine-based agonist, thiaoxoguanosine agonist (7-t hia-8-oxoguanosine-based agonists) The amine group (NH2) and carbamate groups, which are the active sites of agonists for leukocyte-like receptors 7 and 8, (carbamate), disulfide, hydrazine azine, ester, peptide, azide It can synthesize bonds such as β-glucuronide and β-glucuronide. Antibodies, antibody fragments, antibody analogs, etc. can be produced by the reaction. Toll-like receptor 7 Alternatively, the agonist and the antibody may form a linker that is cleaved by various external or internal environmental stimuli. A representative linker is shown in FIG. 1. A conjugate of an antibody and a Toll-like receptor 7 or 8 agonist, The function of the drug is temporarily inhibited and then cut off under certain conditions. To confirm whether a conjugate that activates the toll-like receptor 7 or We produced conjugates linked to antibodies via various bonds using resiquimod, one of the 8-agonist drugs. Ta.

[0063] 10.1. Toll-like receptor 7 or 8 action with protease-cleavable bonds Synthesis of drug-antibody conjugates Antibody and Toll-like receptor 7 can be cleaved by protease Or to synthesize a conjugate in which the agonist is linked by a peptide bond, For this purpose, the method of the following reaction formula 10 or reaction formula 11 was used. As a specific example of reaction formula 10, Cathepsin B cleavage is induced by the sulfhydryl bonds. To synthesize a conjugate of resiquimod with an antibody containing a aryl group, a linker was first added to the activation site. More specifically, resiquimod (10 mg) and Mal were synthesized. -Val-Cit-PAB-PNP (47.2 mg) was dissolved in 2 ml of dimethylformamide ( N,N-Dimethylformamide (DMF) and then dissolved. Then, N,N-diisopropylethylamine thylamine;DIPEA) (6.7 μl) and hydroxybenzotriazole (1-Hydroxybenzotriazole hydrate;1-HOBt)( 315 μg) was added. Then, the mixture was stirred at room temperature for 16 hours to prepare a mixture. The mixture was analyzed by thin-layer chromatography. hy) and cathepsin B activates Toll-like receptor 7 or 8 agonists. He obtained resiquimod, a white powder that dynamically restores function to the area. In addition, in order to reduce the antibody, anti-VEGFR2 antibody, a type of antibody, and Tris (Chloroethyl)phosphate (Tris(2-carboxyethyl)phosphi ne;TCEP) was added to 1 mM EDTA solution (pH 7.4) and reacted at 37°C for 1 hour. Next, after lowering the temperature to 22°C, dehydroascorbic acid (Dehydroa Add scorbic acid to adjust the pH to 6.5, react for 1 hour, and then Cathepsin B then inhibits the activity of Toll-like receptor 7 or 8 agonists. The resiquimod conjugate, which shows kinetic recovery of the function of the ligation site, was analyzed using 1M Tris buffer. After adjusting the pH to 7.4, the antibody was added and reacted at 22°C for 30 minutes to form the conjugate. The produced and unreacted material was removed by gel electrophoresis using a PD-10 column.

[0064] [ka]

[0065] [ka]

[0066] 10.2. Toll-like receptors with bonds cleaved by β-glucuronidase 7 or Synthesis of agonist-antibody conjugates So that it can be cleaved by β-glucuronidase , antibodies and Toll-like receptor 7 or 8 agonists were used to treat β-glucuronidine (β-Glucuronidine To synthesize the conjugate linked by an ide bond, the method shown in the following reaction scheme 12 was used. Next, to reduce the antibody, we mixed anti-VEGFR2 antibody, a type of antibody, with triclosan. S(chloroethyl)phosphate was added to 1 mM EDTA solution (pH 7.4) and incubated for 1 h. The reaction was carried out at 7°C. Next, the temperature was lowered to 22°C, and dehydroascorbic acid was added. The pH was adjusted to 6.5 and the reaction was allowed to proceed for 1 hour for further oxidation. Toll-like receptor 7 or 8 bound to the nidin conjugate, i.e., inactivated The reaction of one of the agonists, resiquimod, with sulfhydryl groups on partially reduced antibodies was investigated. More specifically, the pH was adjusted to 7.4 using 1M Tris buffer. The antibody and β-glucuronidine-conjugated resiquimod were mixed and incubated at 22°C for 30 The conjugate was produced by reacting for 1 min, and the unreacted material was separated by gel electrophoresis using a PD-10 column. was removed by law.

[0067] [ka]

[0068] 10.3. Toll-like receptor 7 or 8 agonists and antagonists with reducing agent-cleavable bonds Body Composition The antibody and the Toll-like receptor are linked together so that they can be cleaved by a reducing agent. Conjugates in which receptor 7 or 8 agonists are linked by disulfide bonds To synthesize this compound, the method shown in the following reaction scheme 13 was used. l) resiquimod (31.4 mg), a toll-like receptor 7 or 8 agonist, and Pyridine (100 μl) was added and dissolved in the solution, and anti-SIRP, a type of antibody, was added. Add α antibody (90 mg) and maleimidocaproyl carbonate disulfide NHS caprylamide gel electrophoresis. Maleimidocaproyl-carbonate-dis sulfide-NHS carbonate ester (90 mg) is added. A dichloromethane (1 ml) solution of 100 ml of 100% ethanol was slowly added dropwise. The mixture was then stirred at 4°C for 16 hours. Distilled water was added to the mixture to form a water and dichloromethane layer. After separation, sodium sulfate was added to the separated dichloromethane layer and reacted for 16 hours. The remaining solution was then purified using a silica gel column to remove the remaining water. White powder of antibody-bound resiquimod was obtained.

[0069] [ka]

[0070] 10.4. Toll-like receptor 7 or 8 agonists and antibodies with bonds that are cleaved under acidic conditions Synthesis of conjugates Antibodies and toll-like receptor 7 or 8 agonists are hydrolyzed so that they can be cleaved under acidic conditions in cells. To synthesize the conjugate linked by a rhazine bond, the method shown in the following reaction scheme 14 was used. In detail, dichloromethane (3 ml) was diluted with one of the Toll-like receptor 7 or 8 agonists, Receptor 1, The antibody was dissolved in a solution containing siquimod (31.4 mg) and pyridine (100 μl). Anti-SIRPα antibody (10 μg), a type of SIRPα antibody, was added, and then maleimide capped. Maleimidoc aproyl-hydroxyethyl hydrazine-NHS carbon A solution of 1 ml of dichloromethane containing 90 mg of ethyl ester was The mixture was then stirred at 4°C for 16 hours to produce a mixture. Distilled water was added to the mixture to separate it into water and dichloromethane layers, and the separated dichloromethane was Sodium sulfate was added to the layer and allowed to react for 16 hours to remove remaining water. The resulting solution was purified using a silica gel column to obtain the white powder of antibody-bound resiquimod. I got it.

[0071] [ka]

[0072] Example 11: Synthesis of conjugates of toll-like receptor 7 or 8 agonists and anticancer drugs Diverse Toll-like receptor 7 or 8 agonists (imidazoquinoline-based agonists) combined with anticancer drugs drugs, hydroxyadenine agonists, pteridone agonists, aminopyrimidine agonists, Drugs that bind to Toll-like receptor 7 or The amine group (NH2) which is the active site of the 8-agonist is linked to the carbamate, disulfide, It can synthesize bonds such as hydrazine, ester, peptide, azide, and β-glucuronidine. It can be produced in reaction with a variety of anticancer drugs. Toll-like receptor 7 or 8 agonists The anticancer drug can be linked to the cytoplasmic reticulum by a linker that can be broken by various external or internal environmental stimuli. The following describes a conjugate of an anticancer drug and a Toll-like receptor 7 or 8 agonist, The function of receptor 7 or 8 agonists is temporarily inhibited, followed by To confirm whether a conjugate that is cleaved and activated under specific conditions is actually produced, We used resiquimod, a drug that acts on receptor-like receptors 7 and 8, to bind anticancer drugs in a variety of ways. A ligated conjugate was prepared.

[0073] 11.1. Anticancer drugs containing amine groups with reducing agent-cleavable bonds and Toll-like receptors Synthesis of conjugates with receptor 7 or 8 agonists Antibodies that contain amine groups so that they can be cleaved by a reducing agent. Cancer drugs (chemo-drugs containing amine groups) and Disulfide-linked agonists for receptors 7 and 8 were To synthesize the conjugate, first, an anticancer drug containing an amine group is To achieve this reduction, doxorubicin and tris(chloroform) were used, which are anticancer drugs that contain amine groups. Tris(2-carboxyethyl)phosphine; TCEP) was added to 1 mM EDTA solution (pH 7.4) and reacted at 37°C for 1 hour. Next, after lowering the temperature to 22°C, dehydroascorbic acid (Dehydroascorbic acid Add rbic acid to adjust the pH to 6.5, react for 1 hour, and further oxidize. Then, the linker between the disulfide in the non-oxidized reduced state and the linker containing resiquimod was Induce the reaction. More specifically, adjust the pH to 7.4 using 1M Tris buffer. After that, the oxidized anticancer drug and resiquimod were mixed and reacted at 22°C for 30 minutes to allow binding. The unreacted material was removed by gel electrophoresis using a PD-10 column. Representative types of anticancer drugs that contain an amine group and a bond that can be cleaved by a reducing agent Examples of conjugates of anticancer drugs and Toll-like receptor 7 or 8 agonists, and those cleaved by reducing agents The mechanism by which this occurs is shown in Figure 2.

[0074] 11.2. Amine groups with bonds that are broken by the hydrogen ion concentration (pH) inside the cell Synthesis of conjugates of anticancer drugs containing Toll-like receptor 7 or 8 agonists Anticancer drugs and toll-like receptor 7 or 8 agonists are cleaved in the acidic conditions inside the cells. A conjugate linked to a hydrazine bond was synthesized. More specifically, dichloromethane (3 ml ) was administered resiquimod (31.4 mg), a Toll-like receptor 7 or 8 agonist, and Lysine (100 μl) was added and dissolved in the solution, and arylhydrazine (90 mg) and Doxorubicin (31.4 mg), an anticancer drug that contains an amine group, is added. A solution of dichloromethane (1 ml) was slowly added dropwise. Then, the mixture was stirred at 4° C. for 16 hours. Distilled water was added to the mixture to separate it into a water layer and a dichloromethane layer. After that, sodium sulfate was added to the separated dichloromethane layer and reacted for 16 hours. The remaining water was removed. The remaining solution was then purified using a silica gel column to give a white We obtained a powdered anticancer drug, Resiquimod, which has an amine bond that can be broken under acidic conditions. Examples of conjugates of anticancer drugs containing an isopropyl group with Toll-like receptor 7 or 8 agonists and The mechanism by which this occurs is shown in Figure 3.

[0075] 11.3. Antibodies containing hydroxy groups with bonds that are cleaved by β-glucuronidase Synthesis of conjugates of cancer drugs with Toll-like receptor 7 or 8 agonists So that it can be cleaved by β-glucuronidase Anticancer drugs and Toll-like receptor 7 or 8 agonists are β-glucuronidine (β-Glucuronidine) More specifically, the conjugate was synthesized by combining dichloromethane (3 ml ) was administered resiquimod (31.4 mg), a Toll-like receptor 7 or 8 agonist, and Lysine (100 μl) was added and dissolved in the solution, and nitrophenyl β-glucuronide (4- Nitrophenyl β-D-glucuronide) (90 mg) and hydrochloride A diluent containing paclitaxel (31.4 mg), an anticancer drug containing an oxy group. A solution of chloromethane (1 ml) was slowly added dropwise. The mixture was then stirred at 4°C for 16 hours. Distilled water was added to the mixture to separate it into a water layer and a dichloromethane layer. After that, sodium sulfate was added to the separated dichloromethane layer and reacted for 16 hours. The remaining solution was then purified using a silica gel column to give a white powder. We obtained resiquimod, which is a compound containing the anticancer drug hydroxyl group. Anticancer drugs with β-glucuronidase-cleavable bonds and Toll-like receptors Examples of conjugates with agonists and the mechanism of cleavage are shown in FIG.

[0076] 11.4. Anticancer drugs containing hydroxy groups with protease-cleavable bonds Synthesis of conjugates with Toll-like receptor 7 or 8 agonists Anticancer drugs and Toll-like receptors that can be cleaved by proteases To synthesize a conjugate in which an agonist is linked to an antibody via a peptide bond, first Paclitaxel, an anticancer drug that contains a hydroxyl group, was used to reduce the Tris(chloroethyl)phosphate hine;TCEP) was added to 1 mM EDTA solution (pH 7.4) and incubated at 37°C for 1 hour. After the temperature was lowered to 22°C, dehydroascorbic acid was added. Add oascorbic acid to adjust the pH to 6.5, and react for 1 hour. The oxidized anticancer drug was phenyl phosphate. sphate (100 mg), followed by one of the Toll-like receptor 7 or 8 agonists. In more detail, the reaction with resiquimod, which is one of the markers of oxidative stress, was induced using 1M Tris buffer. After adjusting the pH to 7.4, the oxidized anticancer drug and resiquimod were mixed and incubated at 22°C. The conjugate was produced by reacting for 30 minutes, and the unreacted material was separated by gel electrophoresis using a PD-10 column. Antibodies containing hydroxy groups with bonds that can be cleaved by proteases were removed by electrophoresis. Examples of conjugates of cancer drugs with Toll-like receptor 7 or 8 agonists and the mechanism of cleavage are shown in Figure 1. 5.

[0077] Example 12: Synthesis of Toll-like receptor 7 or 8 agonist and protein-based drug conjugates Growth Protein-based drugs, i.e., various functional drugs conjugated with proteins, Toll-like receptor 7 or 8 agonists (imidazoquinoline agonists, hydroxyadenine agonists) Drugs for treating rheumatoid arthritis, pteridones, aminopyrimidines, benzazepines, thiazolam Oxoguanosine agonists, etc., are the activation sites of Toll-like receptor 7 or 8 agonists. The amine group (NH2) and carbamates, disulfides, hydrazines, esters, and peptides A variety of protein-based conjugates can be synthesized, including tide, azide, and β-glucuronidine. Drugs can be produced by reacting Toll-like receptor 7 or 8 agonists with proteins The base drug may be linked by a linker that is cleavable by a variety of external or internal environmental stimuli. Below, we present a conjugate of a protein-based drug with a Toll-like receptor 7 or 8 agonist. Therefore, the function of Toll-like receptor 7 or 8 agonists is temporarily inhibited. Then, it is confirmed whether a conjugate that is cleaved and activated under specific conditions is actually produced. To investigate the mechanism of action of resiquimod, a Toll-like receptor 7 or 8 agonist, Conjugates were prepared by linking the base drug with various bonds. The linker and linking mechanism are shown in FIG.

[0078] 12.1. Protein vectors with amine-based reactive bonds that can be cleaved by reducing agents Synthesis of conjugates of toll-like receptor 7 or 8 agonists with various drugs Protein-based drugs and Toll-like receptors 7 or 8 can be cleaved by reducing agents To synthesize conjugates in which the active agent is linked via a disulfide bond, the NHS ester reaction is used. More specifically, NHS ester (3-Mercaptoprop) was used at a temperature of 22°C. yl-N-hydroxysuccinimide ester) to dehydroascorbic acid Dehydroascorbic acid was added to adjust the pH to 6.5. After that, the reaction was continued for 1 hour to produce a conjugate. Then, 1M Tris buffer was added and After further raising the pH to 7.4, the conjugate, a protein-based drug, Protein antigen OVA (ovalbumine) and resiquimod were mixed and incubated at 22°C for 3 The conjugate was produced by reacting for 10 minutes. The unreacted material was separated by gel electrophoresis using a PD-10 column. Protein-based drugs with bonds that are cleaved by reducing agents were removed by gel electrophoresis. Examples of conjugates with agonists of receptor-like receptors 7 and 8 and the mechanism of cleavage are shown in FIG. Ta.

[0079] 12.2. Cleavage due to the hydrogen ion concentration (pH) inside the cell via amine group reaction Conjugates of protein-based drugs with Toll-like receptor 7 or 8 agonists Growth The hydrogen ion concentration inside the cell, i.e., the protein base that can be cleaved under acidic conditions, Synthesis of conjugates in which a toll-like receptor 7 or 8 agonist is linked via a hydrazine bond To achieve this, the NHS ester reaction was used. More specifically, the NHS ester (3-Mer captopropyl-N-hydroxysuccinimide ester) Aryl hydrazine was dissolved in dichloromethane (3 ml) containing pyridine (100 μl). Dichloromethane (1 ml) containing 90 mg of ethanol was slowly added dropwise. The mixture was then added with a protein antigen, OVA (31.4 mg) and resiquimod (31.4 mg) were added and stirred at 4°C for 16 hours to synthesize the conjugate. The synthesized conjugate was then purified using a silica gel column to obtain a white powder protein. The drug is bound to the substrate, resiquimod. It is broken down by the hydrogen ion concentration inside the cell. Binding of protein-based drugs with Toll-like receptor 7 or 8 agonists with targeted binding An example of the polypeptide and the mechanism by which it is cleaved are shown in FIG.

[0080] 12.3. Having a bond that is cleaved by β-glucuronidase via an amine group reaction Synthesis of protein-based drug conjugates with Toll-like receptor 7 or 8 agonists Protein-based drugs and Toll-like receptors that can be cleaved by β-glucuronidase To synthesize conjugates in which receptor 7 or 8 agonists are linked via β-glucuronide, NHS An ester reaction was used. More specifically, resiquimod (31.4 mg), NHS ester ( 3-Mercaptopropyl-N-hydroxysuccinimide Nitrogen was dissolved in dichloromethane (3 ml) containing tert-butyl ether and pyridine (100 μl). Phenyl β-glucuronide (4-Nitrophenyl β-D-glucuroni de) (90 mg) was added dropwise slowly to dichloromethane (1 ml). Then, the mixture was stirred at 4° C. for 16 hours to produce a mixture, and the mixture was added with a protein base. The protein antigen OVA (31.4 mg), one of the drugs, was added and stirred at 4°C for 16 hours. The mixture was stirred to synthesize the conjugate. Next, after adjusting the temperature to room temperature, distilled water was added to synthesize the conjugate. The dichloromethane layer was separated. Sodium sulfate was added to the separated dichloromethane layer, and the mixture was diluted with 16 The reaction was continued for 1 hour to remove the remaining water, and the remaining solution was purified using a silica gel column. To obtain the white powder protein-based drug conjugate resiquimod. Protein-based drugs with cleavable bonds in Toll-like receptor 7 or Examples of conjugates with agonists and the mechanism of cleavage are shown in FIG.

[0081] 12.4. Proteins with sulfhydryl-mediated bonds that are cleaved by reducing agents Synthesis of conjugates of protein-based drugs with Toll-like receptor 7 or 8 agonists Protein-based drugs and Toll-like receptors 7 or 8 can be cleaved by reducing agents To synthesize conjugates in which the agonist is linked by a disulfide bond, maleimide More specifically, a maleimide polyethylene glycol (PEG) reaction was used at a temperature of 22°C. Cole (maleimide PEG) with dehydroascorbic acid (Dehydroas corbic acid) was added to adjust the pH to 6.5, and the mixture was allowed to react for 1 hour. The conjugates were then prepared. They were then mixed with conjugates containing disulfides in the reduced state that were not oxidized. The reaction between the protein antigen OVA (31.4 mg), a protein-based drug, and After that, the pH was adjusted to 9.0, and the protein-based ligase bound to the conjugate was added. The drug and resiquimod (31.4 mg) were mixed and reacted at 22°C for 30 minutes. The non-reactive material was removed by gel electrophoresis using a PD-10 column. Protein-based drugs with reducing agent-cleavable bonds via Toll-like receptors Examples of conjugates with 7 or 8 agonists and the mechanism of cleavage are shown in FIG.

[0082] 12.5. Sulfhydryl reaction is mediated by the hydrogen ion concentration (pH) inside the cell. Binding of protein-based drugs with cleavable bonds to Toll-like receptor 7 or 8 agonists combination synthesis The hydrogen ion concentration inside the cell, i.e., the protein base that can be cleaved under acidic conditions, Synthesis of conjugates in which a toll-like receptor 7 or 8 agonist is linked via a hydrazine bond To achieve this, the maleimide reaction was used. More specifically, maleimide polyethylene glycol Dichloromethane (maleimide PEG) and pyridine (100 μl) were added. Methane (3 ml) to which arylhydrazine (90 mg) was added was mixed in dichloromethane (1 ml l) was slowly added dropwise to the mixture. Then, one of the protein-based drugs was added to the mixture. Protein antigens OVA (31.4 mg) and resiquimod (31.4 mg) were added, and 4 The conjugate was synthesized by stirring at 50° C. for 16 hours. The white powder of protein-based drug-conjugated resiquimod was obtained by purifying it using ruthenium. Protein-based drugs and receptors that have bonds that are broken by the hydrogen ion concentration inside the cell. Examples of conjugates with agonists of receptor-like receptors 7 and 8 and the mechanism of cleavage are shown in FIG. did.

[0083] 12.6. Bonds cleaved by β-glucuronidase via sulfhydryl reactions Synthesis of conjugates of protein-based drugs with Toll-like receptor 7 or 8 agonists Protein-based drugs and Toll-like receptors that can be cleaved by β-glucuronidase To synthesize conjugates in which receptor 7 or 8 agonists were linked with β-glucuronides, More specifically, maleimide polyethylene glycol (maleimide polyethylene glycol) was used. de PEG) and pyridine (100 μl) in dichloromethane (3 ml). Nitrophenyl β-glucuronide (4-Nitrophenyl β-D-glucur Dichloromethane (1 ml) containing 100 g of ethyl phthalate (90 mg) was slowly added dropwise. The pH was then adjusted to 9.0, and the mixture was stirred at 4°C for 16 hours to produce a mixture. The mixture was mixed with the protein antigen OVA ( The mixture was stirred at 4°C for 16 hours to obtain a 31.4 mg solution. Next, after adjusting the temperature to room temperature, distilled water was added and the mixture was mixed with dichloromethane. After separation into a dichloromethane layer, sodium sulfate was added to the separated dichloromethane layer and reacted for 16 hours. The remaining solution was then purified using a silica gel column. The researchers obtained resiquimod, a white powder of protein-based drug conjugate. Protein-based drugs with cleavable bonds toll-like receptor 7 or Examples of 8-agonist conjugates and the mechanism of cleavage are shown in FIG.

[0084] Example 13: Synthesis of conjugates of toll-like receptor 7 or 8 agonists and sensitizers 13.1. Sensitizers with reducing agent-cleavable bonds and Toll-like receptor 7 or 8 agonists Synthesis of conjugates with drugs Toll-like receptor 7 and sensitizers that can be cleaved by reducing agents Or, to synthesize a conjugate in which the eight agonists are linked by disulfide bonds, a photoamplification reaction is first performed. Reduces photosensitizer photofrin To do this, the sensitizer and tris(chloroethyl)phosphate were dissolved in 1 mM EDTA solution (pH 7.4 ) and reacted at 37°C for 1 hour. Then, the temperature was lowered to 22°C, and the dehydrogenase was added. Corbic acid was added to adjust the pH to 6.5, and the reaction was allowed to proceed for 1 hour for further oxidation. We then induced a reaction between the non-oxidized, reduced form of photofrin and resiquimod. More specifically, the pH was adjusted to 7.4 using 1M Tris buffer, and then oxidized. The sensitizer and resiquimod were mixed and reacted at 22°C for 30 minutes to produce a conjugate. The unwanted substances were removed by gel electrophoresis using a PD-10 column. Conjugates of sensitizers with toll-like receptor 7 or 8 agonists that have a bond that can be cleaved by a sensitizer Examples and the mechanism of cleavage are shown in FIG.

[0085] 13.2. Carboxylic Acids with Bonds That Are Broken by the Hydrogen Ion Concentration (pH) Within a Cell Synthesis of conjugates of silyl-containing sensitizers with toll-like receptor 7 or 8 agonists The hydrogen ion concentration inside the cell is low, i.e., the sensitizer and tol are cleaved under acidic conditions. We synthesized conjugates in which receptor 7 or 8 agonists were linked via hydrazine bonds. The study consisted of administering 3 ml of dichloromethane containing resiquimostat, a Toll-like receptor 7 or 8 agonist. Add arylhydride (31.4 mg) and pyridine (100 μl) to the solution and dissolve. Slowly add dropwise a solution of tetracycline (90 mg) in dichloromethane (1 ml). Then, the mixture was stirred at 4°C for 16 hours to prepare a mixture. Photofrin, one of the agents, was added and reacted at 22°C for 30 minutes to produce the conjugate. Distilled water was added to separate the water and dichloromethane layers, and the separated dichloromethane layer was Sodium sulfate was added to the mixture and reacted for 16 hours to remove the remaining water. The solution was purified using a silica gel column to obtain resiquimod bound to the sensitizer as a white powder. The sensitizer containing a carboxyl group with a bond that is cleaved under acidic conditions and the toll-like receptor Examples of conjugates with agonists 7 or 8 and the mechanism of cleavage are shown in FIG.

[0086] 13.3. Contains a carboxyl group with a bond that is cleaved by β-glucuronidase Synthesis of conjugates of sensitizers and toll-like receptor 7 or 8 agonists Sensitizers and toll-like receptor 7 or 8 agonists that can be cleaved by β-glucuronidase The drug is linked to a conjugate via a β-glucuronide bond. More specifically, the toll-like receptor 7 or 8 agonist was dissolved in dichloromethane (3 ml). One of them, resiquimod (31.4 mg) and pyridine (100 μl) were added and dissolved. Nitrophenyl β-glucuronide (90 mg) was added to the solution in dichloromethane. (1 ml) solution was slowly added dropwise. The mixture was then stirred at 4° C. for 16 hours. The mixture was added with photofrin, a photosensitizer, and incubated at 22°C. The reaction was carried out for 30 minutes to produce a conjugate, and distilled water was added to separate the mixture into water and dichloromethane layers. After that, sodium sulfate was added to the separated dichloromethane layer and reacted for 16 hours. The remaining solution was then purified using a silica gel column to obtain a white powder. We obtained resiquimod with an anticancer drug attached. The bond is cleaved by β-glucuronidase. Examples of conjugates of carboxyl-containing sensitizers and Toll-like receptor 7 or 8 agonists having the formula and the mechanism of cleavage is shown in FIG.

[0087] Example 14: Antibodies and Resins Containing Carboxyl Groups with Hydrolytically Cleavable Bonds Kimodo combination 14.1. Antibodies and Residues Containing Carboxyl Groups with Hydrolytically Cleavable Bonds Modo conjugate synthesis Binding of resiquimod to antibodies containing a carboxyl group with a hydrolytically cleavable bond. To synthesize the body, the active site is first hydrolyzed. We have synthesized Toll-like receptor 7 or 8 agonists that dynamically restore the function of , resiquimod (10 mg) and maleimide-polyethyl glycol-hydroxysuccinimide Maleimide-dPEG4-NHS ester (19.6 mg) After adding dichloromethane (DCM) (2 ml), Then, triethylamine (8.8 μL) was added. Then, the mixture was stirred at room temperature for 16 hours. A mixture is prepared, and the mixture is subjected to thin-layer chromatography. The white powder was purified using omatography, and the function of the activated site was confirmed by hydrolysis. We obtained resiquimod, which dynamically restores the activity of the antibody. To achieve this, anti-VEGFR2 antibody (4 mg) was diluted with 1 mM diethylenetriamine (Di 1M borate b containing ethylenetriamine (DTPA) Next, the cells were dispersed in 120 μM tris(2-carboxyethylidene phosphate buffer. l) Phosphine (TCEP) was added and reduced by stirring at 37° C. for 1 hour. The reduced antibody and resiquimod (210 μg) were then mixed and stirred at 4° C. for 1 hour. Then, zeba TM Using a desalting column, The antibody-resiquimod conjugate (AAC-001) was purified by centrifugation at 00×g for 2 min. .

[0088] 14.2. Antibodies and Residues Containing Carboxyl Groups with Hydrolytically Cleavable Bonds Confirmation of anticancer effects of Modo conjugates The antibody-resiquimod conjugate (AAC-001) was synthesized in the same manner as in Example 14.1. To confirm the anticancer effect, we used B16OVA melanoma. The results were evaluated. 5 The B16OVA cancer cells were placed on the back of the mouse. Subcutaneous injection into the 3rd place, and then every 3 days from 3 days later The drugs were injected directly into the cancer tissue four times. Anti-VEGFR2 complex, anti-VEGFR2 with hydrolytically cleavable bond A conjugate of GFR2 antibody and resiquimod (AAC-001) was used. Then, the cancer growth The extent of the disease was confirmed by measuring the size of the tumor, and the survival rate was also measured. The results were repeated at least three times and are shown as the mean ± standard deviation. Statistical significance was determined by the The results were confirmed by ent's t-test, and statistical significance was determined when P<0.05. The results are shown in Figures 16 and 17.

[0089] As shown in Figures 16 and 17, the AAC-001 conjugate inhibited the cytotoxicity of resiquimod alone. (R848), as well as the combined administration of anti-VEGFR2 and resiquimod (R8 48+anti-VEGFR2), not only did it significantly suppress cancer growth, , and was confirmed to increase survival rate.

[0090] 14.3. Antibodies and Residues Containing Carboxyl Groups with Hydrolytically Cleavable Bonds Confirmation of inflammatory cytokine levels in blood of Modo-bound The antibody-resiquimod conjugate (AAC-001) synthesized in the same manner as in Example 14.1 was To confirm the effect of AAC-001 conjugate on the secretion of intracellular inflammatory cytokines, After intravenous injection of either resiquimod or resiquimod into mice, blood samples were taken at 0, 1, 2, and 4 hours. The collected blood samples were centrifuged at 10,000 × g for 20 minutes to separate the serum, The presence of IL-6 in serum was measured using the Invitrogen mouse IL-6 ELISA kit. The concentration of IL-6, a representative inflammatory cytokine present in the blood, was measured. The results are shown in Figure 18. As shown in.

[0091] As shown in Figure 18, in the case of resiquimod, as soon as it was injected into the body, it caused inflammatory Cytokine storm is induced by promoting the secretion of cytokines. It was confirmed that the amount of inflammatory cytokines was not changed in the case of the conjugate.

[0092] Based on the above results, the conjugate of the antibody of the present invention and the toll-like receptor 7 or 8 agonist, i.e. The antibody binds to a cleavable linker at the activation site of the Toll-like receptor 7 or 8 agonist. In the early stages of administration, functional drugs such as antibodies act on the Toll-like receptors. 7 or 8 agonists exist in a transiently inactivated state and are secreted into the tumor microenvironment or target cells. Toll-like receptor 7 or 8 agonists are activated by hydrolase cleavage of the linker in the cells By increasing the concentration of riboflavin, it has a secondary immune activation effect, and compared with simple combination administration, it has a low It was confirmed that the drug showed few side effects and significantly increased therapeutic efficacy.

[0093] Example 15: Antibody containing a carboxyl group with a bond that is cleaved by cathepsin B Resiquimod conjugate 15.1. Antibodies and receptors containing a carboxyl group with a bond that is cleaved by cathepsin B Synthesis of Siquimod Conjugates Carboxyl with a bond cleaved by cathepsin B To synthesize a conjugate of resiquimod with an antibody containing the carboxyl group, the linker is first attached to the activation site. More specifically, resiquimod (10 mg) and Mal-V were synthesized. al-Cit-PAB-PNP (47.2 mg) was dissolved in 2 ml of dimethylformamide (N, N-Dimethylformamide (DMF) and then dissolved. , N,N-Diisomethylpropylethylamine ylamine;DIPEA) (6.7 μl) and hydroxybenzotriazole (1 -Hydroxybenzotriazole hydrate;1-HOBt)(31 5 μg) was added. Then, the mixture was stirred at room temperature for 16 hours to prepare a mixture. The substance was analyzed by thin-layer chromatography. ) and purified to determine the activation site of Toll-like receptor 7 or 8 agonists by cathepsin B. We obtained the white powder resiquimod, which dynamically restores the function of the target cell. A conjugate of resiquimod with an antibody having a more cleavable bond (AAC-002) was synthesized. For this purpose, anti-SIRPα antibody (4 mg) was incubated with 1 mM diethylenetriamine (D 1M borate containing diethylenetriamine (DTPA) After dispersion in the buffer, 150 μM tris(2-carboxyethoxy) Add trichloroethylene phosphine (TCEP) and stir at 37°C for 1 hour to reduce the antibody. The reaction then kinetically restored the function of the partially reduced antibody and the activation site. The mixture was mixed with zeb (480 μg) and stirred at 4°C for 1 hour to synthesize the conjugate. a TM Using a desalting column, 2 The mixture was centrifuged for 1 min to separate only AAC-002, which is a conjugate of the antibody and resiquimod. The mechanism of AC-002 binding is shown in FIG.

[0094] 15.2. Antibodies and receptors containing a carboxyl group with a bond that is cleaved by cathepsin B Confirmation of blood inflammatory cytokine levels in combination with Siquimod The antibody-resiquimod conjugate (AAC-002) synthesized in the same manner as in Example 15.1 was To confirm the effect of AAC-002 conjugates on the secretion of intracellular inflammatory cytokines, Conjugated TLR7 / 8a-Blocked active site) After intravenous injection into mice, blood was collected 2 hours later. Serum was separated by centrifugation at 0,000 × g for 20 min and purified using Invitrogen mouse serum. IL-6 ELISA kit was used to measure the inflammatory cytokines present in serum. The concentration of IL-6 was measured. As a control, a group was injected with phosphate buffered saline. (Blank), resiquimod alone injection group (Bare TLR7 / 8a-U unconjugated, and antibodies that are not conjugated to the activation site but to other sites Group injected with the TLR7 / 8a-L conjugate (Conjugated TLR7 / 8a-L The results are shown in Figure 20.

[0095] As shown in FIG. 20, administration of resiquimod alone or with antibodies In the case of antibody-resiquimod conjugates that bind to other sites rather than to the IL-1 site, the active site is not present. It induces unnecessary stimuli and significantly increases the levels of inflammatory cytokines in the blood at the initial stage of injection. On the other hand, in the case of the complex in which the antibody binds to the activation site, it was confirmed that the antibody inhibits the inflammatory cycle. It was confirmed that the amount of tocainine did not change.

[0096] Example 16: Protein antigens with bonds cleaved by cathepsin B and resiquimod A combination of Protein-based sulfhydryl-containing bonds that are cleaved by cathepsin B To synthesize a conjugate of resiquimod with a protein antigen, one of the drugs, We synthesized resiquimod with a linker attached to the ligation site. ) and Mal-Val-Cit-PAB-PNP (47.2 mg) in 2 ml of dimethyl After adding to formamide (N,N-Dimethylformamide; DMF), After that, N,N-diisomethylpropylethylamine (N,N-Diiso propylethylamine;DIPEA) (6.7 μl) and hydroxybenzoate 1-Hydroxybenzotriazole hydrate;1 -HOBt) (315 μg) was added. Then, the mixture was stirred at room temperature for 16 hours to produce a mixture. The OVA protein antigen (10 mg) was added to the mixture and reacted to obtain the binding. After preparation of the body, thin-layer chromatography (Thin-layer chromato The OVA fragment was purified using a graphy method to obtain OVA fragments having bonds that can be cleaved by cathepsin B. The conjugates of protein antigens and resiquimod were then analyzed by spectroscopic analysis. The size was confirmed using the method described above, and the results are shown in Figure 21.

[0097] As shown in FIG. 21, OVA protein having a bond that is cleaved by cathepsin B It was confirmed that the conjugate of the antigen and resiquimod formed nanoparticles of approximately 99.7 nm in aqueous solution. I acknowledged.

[0098] Based on the above results, the conjugate of the present invention binds to Toll-like receptor 7 or Or, a functional drug is bound to the active site of the agonist, which activates it. When it is temporarily inhibited and then injected into the body, it inhibits inflammatory cytokines. Since it suppresses the secretion of cytokines and non-specific immune reactions and is non-toxic, it significantly reduces side effects. The conjugate can then be absorbed by the physiological environment to reach the desired location. When this happens, Toll-like receptor 7 or 8 agonists and functional drugs separate, enhancing transmission. In addition, only when Toll-like receptor 7 or 8 agonists and functional drugs are used alone, compared with the combined administration of a Toll-like receptor 7 or 8 agonist and a functional drug. It was confirmed that the therapeutic effect can be significantly increased. This is a method in which the active site of a receptor 7 or 8 agonist is linked to a functional drug via a cleavable linker. It is expected that the conjugates of the invention can be effectively used in the treatment of a variety of diseases.

[0099] The above description of the invention is given by way of example only and is not intended to be limiting of the scope of the invention. A person having ordinary skill in the art would be able to make the present invention into a practical application without changing the technical idea or essential features of the present invention. It can be seen that the physical shape can be easily modified. It is to be understood that the described embodiments are illustrative in all respects and not restrictive. It should be. [Industrial Applicability]

[0100] The activation site of the toll-like receptor 7 or 8 agonist of the present invention and the functional drug-cleavable phosphoryl group Carboxy-linked conjugates are temporarily inactive when administered to the body, but then activate tumor Toll-like receptor 7 or 8 agonists and their functional roles in specific conditions in the tumor microenvironment and / or cells Because the drug is separated and activated, side effects such as non-specific hypersensitivity immune reactions can be reduced. Not only can it be used to effectively regulate immune cells by Toll-like receptor 7 or 8 agonists, but it can also be used to effectively regulate immune cells by Toll-like receptor 7 or 8 agonists. At the same time, the functional drug acts, so that the therapeutic effect can be significantly improved. Therefore, it is applicable to all functional drugs that can be administered in combination with Toll-like receptor 7 or 8 agonists. It is possible to use.

Claims

1. A conjugate of a Toll-like receptor 7 or 8 agonist and a functional drug, the conjugate being characterized in that the functional drug and an active site of the Toll-like receptor 7 or 8 agonist are linked via a cleavable linker.

2. The conjugate according to claim 1, characterized in that the functional drug acts primarily, and the inactivated Toll-like receptor 7 or 8 agonist acts secondarily by cleaving the linker at the injection site, tumor microenvironment or target cell, kinetically restoring the active site.

3. The conjugate of claim 1, wherein the cleavable linker is capable of cleaving a chemical bond at the binding site by any one or more factors selected from the group consisting of an enzyme, pH, redox potential, temperature, ultrasound, magnetism, and a light source.

4. 2. The conjugate of claim 1, wherein the cleavable linker is at least one selected from the group consisting of a benzyl elimination-based linker, a trialkyl lock-based linker, a bicine-based linker, an acid labile linker, a lysosomally cleavable peptide, and a cathepsin B cleavable peptide.

5. The conjugate according to claim 4, wherein the cleavable linker comprises one or more bonds selected from the group consisting of carbamate, disulfide, hydrazine, ester, peptide, azide, β-glucuronide, and combinations thereof.

6. 2. The conjugate of claim 1, wherein the cleavable linker further comprises ethylene oxide or ethylene glycol at both or one termini.

7. The conjugate according to claim 1, wherein the Toll-like receptor 7 or 8 agonist is at least one selected from the group consisting of imidazoquinoline-based agonists, hydroxyadenine-based agonists, pteridone-based agonists, 2-aminopyrimidine-based agonists, benzoazepine-based agonists, and thiaoxoguanosine-based agonists.

8. The conjugate according to claim 1, characterized in that the functional drug is one or more selected from the group consisting of an antibody, an antibody fragment, a single-chain antibody, an anticancer drug, an antigen, a cytokine, a protein, a peptide, an amino acid, an oligonucleotide, an enzyme, a lipid, a low molecular weight compound, a glycoprotein, and a targeting ligand.

9. The conjugate of claim 1, wherein the functional drug is at least one selected from the group consisting of an antibody, a protein-based drug, a sensitizer, and an anticancer drug.

10. A composition for regulating immune function, comprising the conjugate according to claim 1 as an active ingredient.

11. The composition for regulating immune function according to claim 10, characterized in that the composition for regulating immune function activates one or more immune cells selected from the group consisting of antigen-presenting cells, B cells, natural killer cells (NK cells) and T cells.

12. The composition for regulating immune function according to claim 10, characterized in that the composition for regulating immune function regulates the function of one or more immune cells selected from the group consisting of Treg (regulatory T cell), MDSC (myeloid derived suppressor cell), and M2 macrophage.

13. The composition for regulating immune function according to claim 10, further comprising any one or more antibodies selected from the group consisting of anti-PD-1, anti-PD-L1, anti-CTLA-4, anti-KIR, anti-LAG3, anti-CD137, anti-OX40, anti-CD276, anti-CD27, anti-GITR, anti-TIM3, anti-41BB, anti-CD226, anti-CD40, anti-CD70, anti-ICOS, anti-CD40L, anti-BTLA, anti-TCR, and anti-TIGIT that target immune checkpoint factors.

14. A pharmaceutical composition for preventing or treating cancer, comprising the conjugate according to claim 1 as an active ingredient.

15. The pharmaceutical composition of claim 14, further comprising a chemical anticancer agent or an immune checkpoint inhibitor.

16. The pharmaceutical composition of claim 14, wherein the pharmaceutical composition inhibits cancer proliferation, metastasis, recurrence, or resistance to anti-cancer treatment.

17. 13. Use of the conjugate of claim 1 for producing a medicament for use in the prevention or treatment of cancer.

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