STAT6 degrader and composition for preventing or treating immune diseases comprising same as active ingredient

Abstract

The present invention relates to a STAT6 degrader and a composition for preventing or treating immune diseases, comprising same as an active ingredient. In the present invention, novel STAT6-PROTAC compounds capable of specifically degrading STAT6 were synthesized, and it was confirmed that the synthesized group of STAT6-PROTAC compounds specifically inhibited STAT6 in dermal fibroblasts. It was also confirmed that the STAT6-PROTAC compounds of the present invention inhibited fibrosis of skin and lung tissues in an animal model exhibiting increased systemic sclerosis disease activity induced by immune sensitization with vimentin. Furthermore, it was confirmed that the STAT6-PROTAC compounds ameliorate systemic sclerosis by inhibiting the expression of dermal fibrotic cytokines and fibrotic factors.

Description

STAT6 decomposer and a composition for the prevention or treatment of immune diseases containing the same as an active ingredient

[0001] The present invention relates to a STAT6 degrading agent and a composition for the prevention or treatment of immune diseases containing the same as an active ingredient.

[0002] Autoimmune diseases refer to conditions in which the body's immune system attacks normal and healthy tissues, organs, or other bodily components. Many autoimmune diseases originate from abnormal immune responses in the human body and induce self-destruction. Famous examples include rheumatoid arthritis, systemic sclerosis, systemic lupus erythematosus, scleroderma, polymyositis, dermatomyositis, anaphylactic purpura, and Sjögren's syndrome. Other diseases such as primary biliary cirrhosis (PBC), chronic active hepatitis, and Hashimoto's thyroiditis are all associated with autoimmune diseases.

[0003] Among these, systemic scleroderma or systemic sclerosis (SSc) is a progressive, debilitating autoimmune disorder also referred to as cutaneous fibrosis, characterized by the excessive deposition of proteins into the extracellular matrix by skin fibroblasts. SSc is caused by activated fibroblasts excessively producing connective tissue components such as collagen, leading to changes in connective tissues—including the skin, blood vessels, gastrointestinal system, lungs, kidneys, muscles, and joints—resulting in functional defects. Depending on the site of onset, SSc is broadly classified into a limited type, where skin hardening occurs only in the hands below the elbows, feet below the knees, and the face; and a diffuse type, where skin hardening appears in more areas, specifically above the elbows and knees, as well as the torso, accompanied by involvement of internal organs such as the kidneys and lungs. Patients with restricted SSc show a 5-year survival rate of over 90%, whereas patients with diffuse SSc have a poor prognosis, exhibiting a 5-year survival rate of only about 50–70%. Patients with diffuse skin disease often show upregulation of unique skin markers, such as type I interferon (IFN)-induced genes. The role that IFN plays in cutaneous fibrosis can be confirmed by recent reports of scleroderma occurring in patients treated with IFN for chronic viral infections. SSc lesions can be characterized by symptoms such as tissue fibrosis, vasculopathy of small blood vessels, and specific autoimmune disease caused by autoantibodies.In particular, autoimmune reactions are a phenomenon in which the immune system forms antibodies against substances present within the human body rather than against external substances due to abnormalities in the immune system, thereby causing various immune diseases. Autoimmune diseases include SSc, systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, and myositis.

[0004] Diseases caused by hypersensitivity reactions are increasing worldwide, but the fundamental causes of these diseases have not been sufficiently identified. Currently, the treatment method for diseases caused by excessive immune responses involves administering immunosuppressants alone or in combination to alleviate or reduce the various symptoms caused by the disease.

[0005] Immunosuppressants refer to various substances used to reduce or block a host's ability to produce antibodies (humoral immune response) or trigger a cellular immune response in response to the action of an antigen. These immunosuppressants can be usefully applied not only in the field of organ transplantation but also for autoimmune diseases such as systemic sclerosis, lupus, and rheumatoid arthritis, as well as for skin hypersensitivity reactions such as atopic dermatitis and allergies. Excellent immunosuppressants must be able to regulate the imbalance of immune responses, ensure safety for the human body, and have a low frequency of disease recurrence during long-term treatment.

[0006] Currently used immunosuppressants include cyclosporine A and FK506. However, as these are naturally derived compounds with complex chemical structures, they are uneconomical due to high raw material procurement costs and carry the risk of various side effects resulting from long-term administration. Therefore, there is an urgent need for the development of new immunosuppressants capable of economical production while exhibiting low toxicity and inducing immune tolerance.

[0007] Meanwhile, a proteolysis targeting chimera (PROTAC) is a heterodimeric molecule in which a ligand of a target protein and a ligand that binds to E3 ligase are linked via a linker. PROTACs bind to both proteins simultaneously, allowing the target protein to approach E3 ligase in very close proximity; through this, E3 ligase recognizes the target protein as a substrate, inducing polyubiquitination and subsequent proteasome degradation. Since specific proteins can be effectively removed from cells using this principle, PROTACs can be used not only as chemical probes to study the function of target proteins but also possess high potential as therapeutic agents for diseases.

[0008] STAT6 (Signal transducer and activator of transcription 6) becomes active through phosphorylation via the mechanisms of IL-4 and IL-13, which play a central role in immune responses. IL-4 and IL-13 bind to cell membrane receptors, activating Janus tyrosine kinases, which then phosphorylate STAT6. This phosphorylated STAT6 forms a dimer, enters the cell nucleus, promotes the transcription of genes involved in immune and anti-inflammatory responses, and activates immune mechanisms within the human body. However, excessively activated STAT6 causes immune-induced allergic diseases such as asthma, atopy, or chronic obstructive pulmonary disease (COPD), and is further linked to autoimmune diseases or cancer. Therefore, inhibiting STAT6 can serve as an effective treatment for immune diseases. However, all STAT6 inhibitors reported to date lack high affinity and selectivity or a clear intracellular mechanism of action.

[0009] Accordingly, the inventors synthesized a novel PROTAC-based compound capable of degrading STAT6 and confirmed its effect in improving systemic sclerosis, thereby completing the present invention.

[0010] The object of the present invention is to provide a compound represented by the following chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0011] [Chemical Formula 1]

[0012]

[0013] In the above chemical formula 1,

[0014] X is a protein targeting moiety, and

[0015] L is the linker, and

[0016] Y is a ubiquitin ligase binding moiety.

[0017] Another object of the present invention is to provide a compound represented by the following chemical formula 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0018] [Chemical Formula 2]

[0019]

[0020] In the above chemical formula 2,

[0021] R1 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a carboxyl group, or an ester group, and

[0022] R2 is either selected from hydrogen or halogen, and

[0023] R3 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, and

[0024] R5 is any one selected from hydrogen, halogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a straight-chain or branched-chain C1-10 alkylamine group, a carboxyl group, or an ester group.

[0025] Another objective of the present invention is to provide a pharmaceutical composition for the prevention or treatment of systemic sclerosis (SSc) comprising the above-mentioned compound as an active ingredient.

[0026] Another objective of the present invention is to provide a method for treating systemic sclerosis (SSc), comprising the step of administering a pharmaceutically effective amount of the above-mentioned compound to an individual.

[0027] In order to achieve the above objective,

[0028] The present invention provides a compound represented by the following chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0029] [Chemical Formula 1]

[0030]

[0031] In the above chemical formula 1,

[0032] X is a protein targeting moiety, and

[0033] L is the linker, and

[0034] Y is a ubiquitin ligase binding moiety.

[0035] In addition, the present invention provides a compound represented by the following chemical formula 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0036] [Chemical Formula 2]

[0037]

[0038] In the above chemical formula 2,

[0039] R1 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a carboxyl group, or an ester group, and

[0040] R2 is either selected from hydrogen or halogen, and

[0041] R3 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, and

[0042] R5 is any one selected from hydrogen, halogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a straight-chain or branched-chain C1-10 alkylamine group, a carboxyl group, or an ester group.

[0043] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of systemic sclerosis (SSc) comprising the above-mentioned compound as an active ingredient.

[0044] In addition, the present invention provides a method for treating systemic sclerosis (SSc), comprising the step of administering a pharmaceutically effective amount of the above-mentioned compound to an individual.

[0045] The present invention synthesized novel STAT6-PROTAC compounds capable of specifically degrading STAT6, and confirmed that the synthesized group of STAT6-PROTAC compounds specifically inhibits STAT6 in skin fibroblasts. Furthermore, it was confirmed that the STAT6-PROTAC compounds of the present invention inhibit fibrosis in skin and lung tissues in an animal model of systemic sclerosis sensitized with vimentin to increase pathological activity. Additionally, by inhibiting the expression of skin fibrosis cytokines and fibrosis factors, the invention improves systemic sclerosis and can be usefully applied in related industries.

[0046] Figure 1 is a figure confirming the STAT6 degradation effect of the novel STAT6-PROTAC compound of the present invention in skin fibroblasts.

[0047] Figure 2 is a Western blot showing the STAT6-specific degradation effect of the novel STAT6-PROTAC compound SD-2744 of the present invention on skin fibroblasts (A: Western blot result, B: quantification of STAT6 result).

[0048] Figure 3 is a Western blot showing the STAT6-specific degradation effect of the novel STAT6-PROTAC compound SD-2782 of the present invention on skin fibroblasts (A: Western blot result, B: quantification of STAT6 result).

[0049] Figure 4 is a Western blot showing the STAT6-specific degradation effect of the novel STAT6-PROTAC compound SD-2877 of the present invention on skin fibroblasts (A: Western blot result, B: quantification of STAT6 result).

[0050] Figure 5 is a Western blot showing the STAT6 degradation effect of the novel STAT6-PROTAC compound SD-2744 of the present invention in spleen cells (A: Western blot result, B: quantification of STAT6 result).

[0051] Figure 6 is a Western blot showing the STAT6 degradation effect of the novel STAT6-PROTAC compound SD-2782 of the present invention in spleen cells (A: Western blot result, B: quantification of STAT6 result).

[0052] Figure 7 is a Western blot showing the STAT6 degradation effect of the novel STAT6-PROTAC compound SD-2877 of the present invention in spleen cells (A: Western blot result, B: quantification of STAT6 result).

[0053] Figure 8 is a figure confirming the half-maximum inhibition concentration of three novel STAT6-PROTAC compounds of the present invention against STAT6.

[0054] Figure 9 illustrates an experimental schedule to confirm the systemic sclerosis improvement effect of the STAT6-PROTAC compound of the present invention.

[0055] Figure 10 shows the tissue fibrosis inhibitory effect of the STAT6-PROTAC compound SD-2744 of the present invention confirmed by H&E and MT staining (A: staining result, B: quantification of staining result).

[0056] Figure 11 shows the tissue fibrosis inhibitory effect of the STAT6-PROTAC compound SD-2782 of the present invention confirmed by H&E and MT staining (A: staining result, B: quantification of staining result).

[0057] Figure 12 shows the tissue fibrosis inhibitory effect of the STAT6-PROTAC compound SD-2877 of the present invention confirmed by H&E and MT staining (A: staining result, B: quantification of staining result).

[0058] Figure 13 is a figure confirming the inhibition of IL-4 and STAT6 by the STAT6-PROTAC compound SD-2744 of the present invention by immunohistochemical staining (A: staining result, B: quantification of staining result).

[0059] Figure 14 is a figure confirming the inhibition of IL-4 and STAT6 by the STAT6-PROTAC compound SD-2877 of the present invention by immunohistochemical staining (A: staining result, B: quantification of staining result).

[0060] Figure 15 is a figure confirming the STAT6 degradation effect of the STAT6-PROTAC compound SD-2744 of the present invention in lung tissue by immunohistochemical staining (A: staining result, B: quantification of staining result).

[0061] Figure 16 is a figure confirming the STAT6 degradation effect of the STAT6-PROTAC compound SD-2782 of the present invention in lung tissue by immunohistochemical staining (A: staining result, B: quantification of staining result).

[0062] Figure 17 is a figure confirming the STAT6 degradation effect of the STAT6-PROTAC compound SD-2877 of the present invention in lung tissue by immunohistochemical staining (A: staining result, B: quantification of staining result).

[0063] Figure 18 is a Western blot analysis of the fibrosis factor inhibition and STAT6 degradation effects of the STAT6-PROTAC compound SD-2744 of the present invention in lung tissue (A: Western blot analysis results, B: quantification of analysis results).

[0064] Figure 19 is a Western blot analysis of the fibrosis factor inhibition and STAT6 degradation effects of the STAT6-PROTAC compound SD-2877 of the present invention in lung tissue (A: Western blot analysis results, B: quantification of analysis results).

[0065] Embodiments of the present invention will be described in detail below with reference to the attached drawings. In the following description, detailed descriptions of technologies well known to those skilled in the art may be omitted. Furthermore, in describing the present invention, detailed descriptions of related known functions or configurations may be omitted if it is determined that such descriptions would unnecessarily obscure the essence of the present invention. Additionally, the terminology used in this specification is used to appropriately express preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the conventions of the field to which the present invention belongs.

[0066] Therefore, the definitions of these terms should be based on the content throughout this specification. Throughout the specification, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0067] The present invention provides a compound represented by the following chemical formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0068]

[0069] In the above chemical formula 1,

[0070] X is a protein targeting moiety, and

[0071] L is the linker, and

[0072] Y is a ubiquitin ligase binding moiety.

[0073] According to one embodiment of the present invention, the protein targeting moiety can target and bind to a protein in vivo, preferably can target and bind to a STAT protein, and more preferably can target and bind to a STAT6 protein.

[0074] According to one embodiment of the present invention, the protein targeting moiety may be a compound represented by the following chemical formula A.

[0075] [Chemical Formula A]

[0076]

[0077] In the above chemical formula A,

[0078] R1 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a carboxyl group, or an ester group, and

[0079] R2 is either selected from hydrogen or halogen, and

[0080] R3 is any one selected from hydrogen, straight-chain or branched-chain C1-10 alkyl groups.

[0081] According to one embodiment of the present invention, the compound represented by the chemical formula A may be one selected from the group of compounds below.

[0082] According to one embodiment of the present invention, the linker may have the structure of the following chemical formula B.

[0083] [Chemical Formula B]

[0084]

[0085] In the above chemical formula B,

[0086] L1 is R4, or does not exist,

[0087] The above R4 is a straight-chain or branched-chain C1-10 alkyl group, and

[0088] L2 is any one selected from a substituted or unsubstituted 3-10 atom heterocyclic group comprising a straight-chain or branched C1-10 alkoxy group, a straight-chain or branched C1-10 alkylamine group, a straight-chain or branched C1-10 alkylamide group, a straight-chain or branched C1-10 alkenyl group, a C3-10 cycloalkyl group, a C3-10 aryloxy group, a C3-10 arylamine group, and one or more heteroatoms selected from N, O, S, Se, and Te.

[0089] L1 may be bonded to a compound of chemical formula A.

[0090] The above linker may use a previously known linker, but is not excluded therefrom, and preferably may be one selected from compounds represented by the following chemical formulas C to G.

[0091] [Chemical Formula C]

[0092]

[0093] In the above chemical formula C,

[0094] n is an integer from 0 to 10.

[0095] [Chemical Formula D]

[0096]

[0097] In the above chemical formula D,

[0098] n is an integer from 0 to 10.

[0099] [Chemical Formula E]

[0100]

[0101] In the above chemical formula E,

[0102] n is an integer from 0 to 10.

[0103] [Chemical Formula F]

[0104]

[0105] In the above chemical formula F,

[0106] n is an integer from 0 to 10.

[0107] [Chemical Formula G]

[0108]

[0109] In the above chemical formula G,

[0110] n is an integer from 0 to 10.

[0111] According to one embodiment of the present invention, the ubiquitin ligase binding moiety may bind to E3 ubiquitin ligase.

[0112] The above ubiquitin ligase binding moiety may be one selected from compounds represented by the following chemical formulas H to L, preferably optionally substituted thalidomide, pomalidomide, lenalidomide, prolinamide, stereoisomers thereof, or derivatives thereof, and more preferably optionally substituted thalidomide or 1-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide as in the following compound.

[0113] [Chemical Formula H]

[0114]

[0115] In the above chemical formula H,

[0116] Z is H2 or O, and

[0117] The above compound is bonded to L2 of chemical formula B.

[0118] [Chemical Formula I]

[0119]

[0120] In the above chemical formula I,

[0121] Z is H2 or O, and

[0122] The above compound is bonded to L2 of chemical formula B.

[0123] [Chemical Formula J]

[0124]

[0125] In the above chemical formula J,

[0126] Z is H2 or O, and

[0127] The above compound is bonded to L2 of chemical formula B.

[0128] [Chemical Formula K]

[0129]

[0130] In the above chemical formula K,

[0131] Z is H2 or O, and

[0132] The above compound is bonded to L2 of chemical formula B.

[0133] [Chemical Formula L]

[0134]

[0135] In the above chemical formula L,

[0136] Z is H2 or O, and

[0137] The above compound is bonded to L2 of chemical formula B.

[0138] The compound of Formula 1 according to the present invention may be one selected from among the compounds represented by Formulas 3 to 37 of Table 1 below;

[0139] Chemical Formula Name Structure 3D-2737 4D-2738 5D-2739 6D-2740 7D-2741 8D-2742 9D-2743 10D-2744 11D-2745 12D-2746 13D-2764 14D-2765 15D-2766 16D-2767 17D-2768 18D-2769 19D-2770 20D-2780 21D-2781 22D-2782 23D-2783 24D-2875 25D-2876 26D-2877 27D-2902 28D-2909 29D-2920 30D-2921 31D-2922 32D-2923 33D-2924 34D-2925 35D-2926 36D-2927 37D-2929

[0140] In addition, the present invention provides a compound represented by the following chemical formula 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[0141] [Chemical Formula 2]

[0142]

[0143] In the above chemical formula 2,

[0144] R1 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a carboxyl group, or an ester group, and

[0145] R2 is either selected from hydrogen or halogen, and

[0146] R3 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, and

[0147] R5 is any one selected from hydrogen, halogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a straight-chain or branched-chain C1-10 alkylamine group, a carboxyl group, or an ester group.

[0148] In the above chemical formula 2, R1 may be any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a carboxyl group, or an ester group, preferably hydrogen or a straight-chain or branched-chain C1-10 alkyl group, and more preferably hydrogen or substituted methyl 2,2-dimethylpropanoate. The substituted methyl 2,2-dimethylpropanoate may have the structure of the following compound.

[0149]

[0150] R2 may be selected from hydrogen or halogen, and preferably may be hydrogen or fluorine (F).

[0151] R3 may be any one selected from hydrogen, straight-chain or branched-chain C1-10 alkyl groups, and preferably may be hydrogen or methyl (Me).

[0152] R5 may be any one selected from hydrogen, halogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a straight-chain or branched-chain C1-10 alkylamine group, a carboxyl group, or an ester group.

[0153] The compound of Formula 2 according to the present invention may be one selected from the group of compounds below, and said compound may target a protein in vivo, preferably may target a STAT protein, and more preferably may target a STAT6 protein.

[0154] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of systemic sclerosis (SSc) comprising the above-mentioned compound as an active ingredient.

[0155] The term “prevention” as used in this invention refers to any act of suppressing the symptoms of a specific disease or delaying its progression through the administration of the composition of this invention.

[0156] The term “treatment” as used in this invention refers to any act of improving or beneficially altering the symptoms of a specific disease through the administration of the composition of this invention.

[0157] The pharmaceutical composition of the present invention may additionally include an adjuvant in addition to the active ingredient. Any adjuvant known in the art may be used without limitation, but, for example, Freund's complete or incomplete adjuvant may be further included to increase the effect.

[0158] The pharmaceutical composition according to the present invention may be prepared in a form in which an active ingredient is incorporated into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients, and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers that can be used in the pharmaceutical composition of the present invention are not limited to these, but may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0159] The pharmaceutical composition of the present invention may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external formulations, suppositories, or sterile injectable solutions, each according to conventional methods.

[0160] When formulating, the product may be prepared using diluents or excipients such as commonly used fillers, volume expanders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules, and such solid dosage forms may be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc., with the active ingredient. In addition, lubricants such as magnesium stearate and talc may also be used in addition to simple excipients. Liquid dosage forms for oral administration include suspensions, liquid formulations, emulsions, and syrups, and may contain various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, in addition to commonly used diluents such as water and liquid paraffin. Preparations for parenteral administration include sterile aqueous solutions, water-insoluble solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Water-insoluble solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Bases for suppositories may include Witepsol, Tween 61, cocoa paste, laurin paste, glycerogelatin, etc.

[0161] The pharmaceutical composition according to the present invention may be administered to an individual by various routes. Any mode of administration may be anticipated, for example, by oral, intravenous, intramuscular, subcutaneous, or intraperitoneal injection.

[0162] The dosage of the pharmaceutical composition according to the present invention is selected by taking into consideration the age, weight, gender, physical condition, etc. of the individual. It is obvious that the concentration of the active ingredient included in the pharmaceutical composition can be selected in various ways depending on the subject, and preferably, it is included in the pharmaceutical composition at a concentration of 0.01 to 5,000 μg / ml. If the concentration is less than 0.01 μg / ml, pharmaceutical activity may not appear, and if it exceeds 5,000 μg / ml, it may exhibit toxicity to the human body.

[0163] According to one embodiment of the present invention, the compound may target STAT6 (Signal transducer and activator of transcription 6), and targeting STAT6 may degrade STAT6.

[0164] According to one embodiment of the present invention, the compound may inhibit tissue fibrosis, and inhibiting tissue fibrosis may reduce skin thickness and reduce the pulmonary fibrosis index.

[0165] According to one embodiment of the present invention, inhibiting tissue fibrosis may involve reducing collagen in the tissue.

[0166] According to one embodiment of the present invention, inhibiting tissue fibrosis may involve inhibiting the expression of a fibrosis cytokine or a fibrosis factor, wherein the fibrosis cytokine may be IL-4 and the fibrosis factor may be α-smooth muscle actin (α-SMA).

[0167] According to one embodiment of the present invention, the compound may reduce the amount of STAT6 protein in the tissue.

[0168] In addition, the present invention provides a method for treating systemic sclerosis (SSc), comprising the step of administering a pharmaceutically effective amount of the above-mentioned compound to an individual.

[0169] The therapeutic method of the present invention comprises administering the recombinant peptide or the recombinant vector to an individual in a therapeutically effective amount. It is preferable to apply a specific therapeutically effective amount for a specific individual differently depending on various factors, including the specific composition (such as the type and degree of the response to be achieved and whether other agents are used in some cases), the individual's age, body weight, general health status, gender and diet, time of administration, route of administration and secretion rate of the composition, duration of treatment, and drugs used together or concurrently with the specific composition, as well as similar factors well known in the pharmaceutical field. The daily dosage is 0.0001 to 100 mg / kg based on the amount of the pharmaceutical composition of the present invention, preferably 0.01 to 100 mg / kg, and may be administered 1 to 6 times a day. However, it is obvious to those skilled in the art that the dosage or administration amount of each active ingredient must be such that it does not contain an excessively high content of each active ingredient to cause side effects. Therefore, it is preferable to determine the effective amount of the composition suitable for the purpose of the present invention by considering the aforementioned matters.

[0170] The above-mentioned individual is applicable to any mammal, and said mammal includes not only humans and primates, but also livestock such as cattle, pigs, sheep, horses, dogs, and cats.

[0171] The compounds of the present invention may be administered to mammals, such as rats, mice, livestock, and humans, by various routes. All modes of administration are expected, for example, orally, rectally or intravenously, intramuscularly, subcutaneously, intrathecally, or intracerebroventricularly.

[0172] The present invention will be explained in more detail below through examples. These examples are merely for the purpose of explaining the present invention more specifically, and it will be obvious to those skilled in the art that the scope of the present invention is not limited to these examples.

[0173] <Preparation Example 1> Preparation of Compound 3

[0174] Compound 3 of the present invention was prepared using the following reaction formula 1.

[0175] [Reaction Equation 1]

[0176]

[0177] 1-1. Preparation of Compound 1 (ethyl 3-(benzylamino)propanoate)

[0178] Triethylamine (0.7 eq) was added to a solution of benzylamine (1 eq) in toluene at room temperature. The reaction solution was cooled to 0 °C, and then ethyl-3-bromopropionate (0.66 eq) was added. The reaction mixture was stirred at room temperature for 3–6 hours. After the reaction was complete, the solvent was evaporated, and the residue was washed with ethyl acetate and water. Subsequently, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex = 1:1 solvent to obtain Compound 1 with a yield of approximately 54%.

[0179] 1 H NMR (600 MHz, DMSO) δ 7.32-7.25 (m, 4H), 7.22-7.17 (m, 1H), 4.03 (q, J = 7.1 Hz, 2H), 3.66 (s, 2H), 2.69 (t, J = 6.9 Hz, 2H), 2.41 (t, J) = 6.8 Hz, 2H), 2.15 (s, 1H), 1.16 (t, J = 7.1 Hz, 3H).

[0180] 1-2. Preparation of compound 2 (tert-butyl (S)-2-(benzyl(3-ethoxy-3-oxopropyl)carbamoyl)pyrrolidine-1-carboxylate)

[0181] Compound 1 (1 eq), (tert-butoxycarbonyl)-L-proline (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex=1:3 solvent to obtain Compound 2 in a yield of approximately 69%.

[0182] 1 H NMR (600 MHz, DMSO) δ 7.39-7.33 (m, 1H), 7.33-7.17 (m, 4H), 4.60-4.40 (m, 3H), 4.05-3.98 (m, 2H), 3.57-3.43 (m, 2H), 3.41-3.34 (m, 1H), 3.31-3.24 (m, 1H), 2.61-2.50 (m, 1H), 2.48-2.37 (m, 1H), 2.30-2.14 (m, 1H), 2.09-2.02 (m, 1H), 1.79-1.64 (m, 2H), 1.31 (d, J = 30.4 Hz, 9H), 1.15 (t, J = 7.1, 5.4 Hz, 3H).

[0183] 1-3. Preparation of compound 3 (3-((S)-N-benzyl-1-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dimethyl butanoyl) pyrrolidine-2-carboxamido)propanoate)

[0184] Compound 2 was treated with 25% TFA / DCM for 1 hour at room temperature. Volatile substances were removed under vacuum, and the residue was used as is for the next reaction step. The crude amine compound (1 eq), N-(tert-Butoxycarbonyl)-L-tert-leucine (1 eq), HATU (1.5 eq), and DIPEA (3 eq) obtained from Compound 2 were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Subsequently, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex = 1:3 solvent to obtain Compound 3 with a yield of approximately 46%.

[0185] 1 H NMR (600 MHz, DMSO) δ 7.40 (d, J = 7.2 Hz, 1H), 7.36 (t, J = 7.7 Hz, 1H), 7.31-7.19 (m, 3H), 6.62-6.50 (m, 1H), 4.81-4.65 (m, 2H), 4.34 (d, J = 15.4 Hz, 1H), 4.19 (d, J = 9.3 Hz, 1H), 4.05-3.97 (m, 2H), 3.84-3.76 (m, 1H), 3.74-3.66 (m, 1H), 3.63-3.55 (m, 1H), 3.54-3.44 (m, 1H), 2.84-2.74 (m, 1H), 2.65-2.56 (m, 1H), 2.26-2.18 (m, 1H), 2.07-1.99 (m, 1H), 1.89-1.62 (m, 2H), 1.36 (d, J = 11.0 Hz, 9H), 1.18-1.11 (m, 3H), 0.96 (d, J = 11.5 Hz, 9H).

[0186] <Preparation Example 2> Preparation of Intermediate 1 (Int.1)

[0187] Intermediate 1 of the present invention was prepared using the following reaction scheme 2.

[0188] [Reaction Equation 2]

[0189]

[0190] 2-1. Preparation of compound a (tert-butyl (E)-3-(4-((diethoxyphosphoryl)methyl)phenyl)acrylate)

[0191] Triethylamine (2 eq) was added to a MeCN solution containing (4-iodo-benzyl)-phosphonic acid diethyl ester (1 eq), Pd(OAc)2 (10 mol%), and triorthotolylphosphine (10%). Tet-butyl acrylate (2 eq) was added to this reaction mixture, and the mixture was heated in a sealed tube at 110 °C for 4 hours. The reaction solvent was evaporated, and the residue was washed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex = 1:1 solvent to obtain compound a with a yield of approximately 96%.

[0192] 1 H NMR (600 MHz, DMSO) δ 7.64 (d, J = 7.9 Hz, 2H), 7.53 (d, J = 16.0 Hz, 1H), 7.33-7.29 (m, 2H), 6.50 (d, J = 15.9 Hz, 1H), 3.99-3.92 (m, 4H), 3.28 (d, J = 21.9 Hz, 2H), 1.49 (s, 9H), 1.17 (t, J = 7.0 Hz, 6H).

[0193] 2-2. Preparation of intermediate 1 (Int.1; (E)-3-(4-((diethoxyphosphoryl)methyl)phenyl)acrylic acid)

[0194] Compound a was treated with 25% TFA / DCM for 1 hour at room temperature. Volatile substances were removed under vacuum, and the resulting crude material Int. 1 was used in the next step without purification.

[0195] <Preparation Example 3> Preparation of Intermediate 2 (Int. 2)

[0196] Intermediate 2 of the present invention was prepared using the following reaction formula 3 or reaction formula 4.

[0197] [Reaction Equation 3]

[0198]

[0199] [Reaction Equation 4]

[0200]

[0201] 3-1. Preparation of Compound b (2-(2,6-dioxopiperidin-3-yl)-5-fluoroisoindoline-1,3-dione)

[0202] 5-fluoroisobenzofuran-1,3-dione (1 eq), 3-aminopiperidine-2,6-dione (1 eq), and NaOAc (2 eq) were added to AcOH and stirred overnight at 110 °C. After evaporating the solvent and adding water, the resulting solution was stirred for 30 minutes. Subsequently, the solid was filtered through a vacuum pump to obtain compound b in a yield of 83%.

[0203] 1 H NMR (600 MHz, DMSO-d6) δ 11.13 (s, 1H), 8.02-7.98 (m, 1H), 7.86-7.82 (m, 1H), 7.74-7.69 (m, 1H), 5.18-5.12 (m, 1H), 2.92-2.84 (m, 1H), 2.63-2.56 (m, 1H), 2.56-2.50 (m, 1H), 2.09-2.02 (m, 1H).

[0204] 3-2. Preparation of Compound bi(tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethyl) carbamate)

[0205] A DMSO solution containing compound b (1 eq), N-Boc-1,2-diaminoethane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. Afterward, the mixture was diluted with water and extracted with ethyl acetate. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound bi in a yield of 28%.

[0206] 1 H NMR (600 MHz, CDCl3) δ 8.18 (s, 1H), 7.58 (d, J = 8.3 Hz, 1H), 6.94-6.91 (m, 1H), 6.74-6.71 (m, 1H), 5.50-5.44 (m, 1H), 4.96-4.93 (m, 1H), 4.93-4.88 (m, 1H), 3.46-3.40 (m, 2H), 3.35-3.30 (m, 2H), 1.67-1.59 (m, 2H), 1.46 (s, 9H).

[0207] 3-3. Compound b.ii (tert-butyl (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)propyl) carbamate) Preparation

[0208] A DMSO solution containing compound b (1 eq), N-Boc-1,3-diaminopropane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. Afterward, the mixture was diluted with water and extracted with ethyl acetate. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound b.ii in a yield of 27%.

[0209] 1H NMR (600 MHz, CDCl3) δ 8.09 (s, 1H), 7.59 (d, J = 8.3 Hz, 1H), 7.00-6.94 (m, 1H), 6.80-6.73 (m, 1H), 5.41-5.33 (m, 1H), 4.97-4.90 (m, 1H), 4.74-4.63 (m, 1H), 3.33-3.20 (m, 4H), 1.81-1.73 (m, 2H), 1.66-1.54 (m, 2H), 1.46 (s, 9H).

[0210] 3-4. Preparation of compound b.iii (tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)butyl)carbamate)

[0211] A DMSO solution containing compound b (1 eq), N-Boc-1,4-diaminobutane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. Afterward, the mixture was diluted with water and extracted with ethyl acetate. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound b.iii in a yield of 30%.

[0212] 1 H NMR (600 MHz, CDCl3) δ 8.55 (s, 1H), 7.53 (d, J = 8.3 Hz, 1H), 6.91-6.82 (m, 1H), 6.71-6.62 (m, 1H), 5.04-4.99 (m, 1H), 4.96-4.91 (m, 1H), 4.72-4.66 (m, 1H), 3.23-3.12 (m, 4H), 1.82-1.70 (m, 1H), 1.69-1.63 (m, 2H), 1.63-1.56 (m, 2H), 1.45 (s, 9H), 1.43-1.40 (m, 1H).

[0213] 3-5. Preparation of compound b.iv (tert-butyl (5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)pentyl) carbamate)

[0214] A DMSO solution containing compound b (1 eq), N-Boc-1,5-diaminopentane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. Afterward, the mixture was diluted with water and extracted with ethyl acetate. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compounds b and iv in a yield of 31%.

[0215] 1 H NMR (600 MHz, CDCl3) δ 8.44 (s, 1H), 7.55 (d, J = 8.3, 1.7 Hz, 1H), 6.93-6.84 (m, 1H), 6.71-6.67 (m, 1H), 4.96-4.91 (m, 1H), 4.90-4.85 (m, 1H), 4.67-4.61 (m, 1H), 3.20-3.10 (m, 4H), 1.81-1.72 (m, 1H), 1.69-1.60 (m, 2H), 1.56-1.48 (m, 2H), 1.48-1.38 (m, 12H).

[0216] 3-6. Preparation of Compound bv (tert-butyl (6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)hexyl) carbamate)

[0217] A DMSO solution containing compound b (1 eq), N-Boc-1,6-diaminohexane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. Afterward, the mixture was diluted with water and extracted with ethyl acetate. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound bv in a yield of 35%.

[0218] 1 H NMR (600 MHz, CDCl3) δ 8.16 (s, 1H), 7.59 (d, J = 8.3 Hz, 1H), 6.96-6.92 (m, 1H), 6.75-6.71 (m, 1H), 4.96-4.91 (m, 1H), 4.77-4.71 (m, 1H), 4.59-4.52 (m, 1H), 3.23-3.18 (m, 2H), 3.16-3.10 (m, 2H), 1.68-1.62 (m, 3H), 1.53-1.47 (m, 2H), 1.47-1.40 (m, 12H), 1.40-1.34 (m, 2H).

[0219] 3-7. Preparation of Compound c (2-(2,6-dioxopiperidin-3-yl)-4-fluoro isoindoline-1,3-dione)

[0220] 4-fluoroisobenzofuran-1,3-dione (1 eq), 3-aminopiperidine-2,6-dione (1 eq), and NaOAc (2 eq) were added to AcOH and stirred overnight at 110 °C. After evaporating the solvent and adding water, the resulting solution was stirred for 30 minutes. Subsequently, the solid was filtered through a vacuum pump to obtain compound c in a yield of 71%.

[0221] 1H NMR (600 MHz, DMSO-d6) δ 11.14 (s, 1H), 7.96-7.91 (m, 1H), 7.78 (d, J = 7.3 Hz, 1H), 7.73 (t, J = 8.8 Hz, 1H), 5.17-5.12 (m, 1H), 2.91-2.84 (m, 1H), 2.63-2.56 (m, 1H), 2.54-2.49 (m, 1H), 2.08-2.02 (m, 1H).

[0222] 3-8. Preparation of Compound ci (tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl) carbamate)

[0223] A DMSO solution containing compound c (1 eq), N-Boc-1,2-diaminoethane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. The mixture was diluted with water and extracted with EA. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound ci in a yield of 46%.

[0224] 1 H NMR (600 MHz, CDCl3) δ 8.26 (s, 1H), 7.50 (t, J = 7.4 Hz, 1H), 7.11 (d, J = 7.1 Hz, 1H), 6.99 (d, J = 8.5 Hz, 1H), 6.40 (t, J = 6.1 Hz, 1H), 4.95-4.88 (m, 2H), 3.45 (q, J = 6.1 Hz, 2H), 3.36 (q, J = 6.1 Hz, 2H), 2.91-2.86 (m, 1H), 2.84-2.71 (m, 2H), 2.14-2.10 (m, 1H), 1.45 (s, 9H).

[0225] 3-9. Compound c.ii (tert-butyl (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl) carbamate) preparation

[0226] A DMSO solution containing compound c (1 eq), N-Boc-1,3-diaminopropane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. The mixture was diluted with water and extracted with EA. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound c.ii in a yield of 44%.

[0227] 1 H NMR (600 MHz, CDCl3) δ 8.54 (s, 1H), 7.50-7.43 (m, 1H), 7.09-7.04 (m, 1H), 6.90-6.84 (m, 1H), 6.32 (t, J = 5.8 Hz, 1H), 4.95-4.89 (m, 1H), 4.77 (t, J = 6.4 Hz, 1H), 3.32 (q, J = 6.0 Hz, 2H), 3.24 (d, J = 7.3 Hz, 2H), 2.87 (d, J = 14.7 Hz, 1H), 2.82-2.69 (m, 2H), 2.14-2.07 (m, 1H), 1.83 (t, J = 7.1 Hz, 2H), 1.44 (s, 9H).

[0228] 3-10. Preparation of Compound c.iii (tert-butyl (4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl) carbamate)

[0229] A DMSO solution containing compound c (1 eq), N-Boc-1,4-diaminobutane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. The mixture was diluted with water and extracted with EA. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound c.iii in a yield of 40%.

[0230] 1 H NMR (600 MHz, CDCl3) δ 8.40 (s, 1H), 7.50-7.45 (m, 1H), 7.10-7.06 (m, 1H), 6.88 (d, J = 8.5 Hz, 1H), 6.24 (t, J = 5.9 Hz, 1H), 4.94-4.89 (m, 1H), 4.63 (s, 1H), 3.29 (d, J = 6.8 Hz, 2H), 3.17 (d, J = 7.2 Hz, 2H), 2.88 (d, J = 14.5 Hz, 1H), 2.83-2.69 (m, 2H), 2.15-2.10 (m, 1H), 1.72-1.65 (m, 2H), 1.63-1.56 (m, 2H), 1.44 (s, 9H).

[0231] 3-11. Preparation of Compound c.iv (tert-butyl (5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentyl) carbamate)

[0232] A DMSO solution containing compound c (1 eq), N-Boc-1,5-diaminopentane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. The mixture was diluted with water and extracted with EA. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compounds c and iv in a yield of 50%.

[0233] 1 H NMR (600 MHz, CDCl3) δ 8.30 (s, 1H), 7.51-7.46 (m, 1H), 7.10-7.06 (m, 1H), 6.89-6.84 (m, 1H), 6.23 (t, J = 5.6 Hz, 1H), 4.94-4.88 (m, 1H), 4.58 (s, 1H), 3.26 (q, J = 5.7 Hz, 2H), 3.13 (d, J = 6.4 Hz, 2H), 2.93-2.84 (m, 1H), 2.84-2.69 (m, 2H), 2.17-2.08 (m, 1H), 1.71-1.65 (m, 2H), 1.57-1.49 (m, 2H), 1.48-1.40 (m, 11H).

[0234] 3-12. Preparation of Compound cv (tert-butyl (6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexyl) carbamate)

[0235] A DMSO solution containing compound c (1 eq), N-Boc-1,6-diaminohexane (1 eq), and DIPEA (2 eq) was microwaved at 130.0°C for 1 hour. The mixture was diluted with water and extracted with EA. Subsequently, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex=1:1 solvent to obtain compound cv in a yield of 40%.

[0236] 1H NMR (600 MHz, CDCl3) δ 8.29 (s, 1H), 7.51-7.47 (m, 1H), 7.10-7.07 (m, 1H), 6.87 (d, J = 8.5 Hz, 1H), 6.23 (t, J = 5.6 Hz, 1H), 4.94-4.90 (m, 1H), 4.55 (s, 1H), 3.29-3.23 (m, 2H), 3.12 (d, J = 6.8 Hz, 2H), 2.91-2.86 (m, 1H), 2.84-2.72 (m, 2H), 2.15-2.11 (m, 1H), 1.69-1.63 (m, 2H), 1.53-1.47 (m, 2H), 1.46-1.40 (m, 11H), 1.40-1.35 (m, 2H).

[0237] 3-13. Preparation of Intermediate Int. 2

[0238] Intermediates Int. 2.bi, Int. 2.b.ii, Int. 2.b.iii, Int. 2.b.iv, Int. 2.bv, Int. 2.ci, Int. 2.c.ii, Int. 2.c.iii, Int. 2.c.iv, and Int. 2.cv were produced by reacting compounds bi, b.ii, b.iii, b.iv, bv, ci, c.ii, c.iii, c.iv, and cv with a 25% TFA / DCM solution at room temperature for 1 hour, respectively, and removing volatile components under reduced pressure; the resulting amine compounds were used directly as crude materials in the next reaction step.

[0239] <Preparation Example 4> Preparation of Compound 4 and Compound 5

[0240] Compounds 4 and 5 of the present invention were prepared using the following reaction scheme 5.

[0241] [Reaction Equation 5]

[0242]

[0243] 4-1. Preparation of compound 4 (ethyl 3-((S)-N-benzyl-1-((S)-2-((E)-3-(4-((diethoxyphosphoryl)methyl)phenyl)acrylamido)-3,3-dimethylbutanoyl)pyrrolidine-2-carboxamido)propanoate)

[0244] Compound 3 was treated with 25% TFA / DCM for 1 hour at room temperature. Volatile substances were removed under vacuum, and the resulting crude was used directly in the next reaction step. Subsequently, the acid crude of Compound 1 (1 eq), the amine crude of Compound 3 (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using a MeOH:MC = 1:15 solvent to obtain Compound 4 with a yield of approximately 73%.

[0245] 1H NMR (600 MHz, DMSO) δ 8.22-8.15 (m, 1H), 7.54 (d, J = 7.5 Hz, 1H), 7.49 (t, J = 7.6 Hz, 2H), 7.44-7.40 (m, 2H), 7.40-7.35 (m, 1H), 7.34-7.31 (m, 1H), 7.30-7.24 (m, 3H), 7.01-6.95 (m, 1H), 4.78-4.73 (m, 1H), 4.70 (d, J = 9.2 Hz, 1H), 4.34 (d, J = 15.4 Hz, 1H), 4.06-3.90 (m, 5H), 3.76-3.62 (m, 2H), 3.57-3.49 (m, 1H), 3.30-3.22 (m, 2H), 2.85-2.78 (m, 1H), 2.65-2.57 (m, 1H), 2.38 (s, 2H), 2.28-2.21 (m, 1H), 2.11-2.03 (m, 1H), 1.99-1.87 (m, 1H), 1.86-1.73 (m, 1H), 1.72-1.65 (m, 1H), 1.20-1.15 (m, 6H), 1.13 (t, J = 7.1 Hz, 3H), 1.03 (d, J = 10.2 Hz, 9H).

[0246] 4-2. 화합물 5a (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0247] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.bi (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5a was obtained in a yield of 20% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0248] 1H NMR (600 MHz, DMSO) δ 11.06 (s, 1H), 8.14 (d, J = 9.0 Hz, 1H), 8.07 (d, J = 35.7 Hz, 1H), 7.58-7.52 (m, 1H), 7.51-7.44 (m, 2H), 7.43-7.19 (m, 8H), 7.14-7.09 (m, 1H), 6.98-6.91 (m, 2H), 6.88-6.82 (m, 1H), 5.06-5.00 (m, 1H), 4.81-4.73 (m, 2H), 4.71-4.65 (m, 1H), 4.26 (d, J = 15.5 Hz, 1H), 3.97-3.89 (m, 5H), 3.88-3.75 (m, 1H), 3.69-3.61 (m, 1H), 3.45-3.38 (m, 1H), 3.29-3.17 (m, 6H), 2.91-2.82 (m, 1H), 2.61-2.53 (m, 1H), 2.52-2.50 (m, 1H), 2.45-2.31 (m, 1H), 2.29-2.18 (m, 1H), 2.11-1.95 (m, 2H), 1.95-1.65 (m, 3H), 1.15 (t, J = 7.0 Hz, 6H), 1.02 (d, J = 15.8 Hz, 9H).

[0249] 4-3. 화합물 5b (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0250] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.b.ii (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5b was obtained in a yield of 15% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0251] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 8.03-7.94 (m, 1H), 7.56-7.52 (m, 1H), 7.49-7.45 (m, 2H), 7.41-7.19 (m, 8H), 7.10-7.05 (m, 1H), 6.97-6.91 (m, 2H), 6.84-6.79 (m, 1H), 5.05-4.99 (m, 1H), 4.81-4.73 (m, 2H), 4.70-4.64 (m, 1H), 4.25 (d, J = 15.4 Hz, 1H), 3.97-3.88 (m, 5H), 3.87-3.77 (m, 1H), 3.68-3.60 (m, 1H), 3.42-3.38 (m, 1H), 3.27-3.20 (m, 2H), 3.19-3.09 (m, J = 8.1 Hz, 4H), 2.90-2.82 (m, 1H), 2.60-2.50 (m, 2H), 2.48-2.34 (m, 1H), 2.28-2.18 (m, 1H), 2.11-2.03 (m, 1H), 2.01-1.95 (m, 1H), 1.94-1.61 (m, 5H), 1.17-1.13 (m, 6H), 1.01 (d, J = 16.6 Hz, 9H).

[0252] 4-4. 화합물 5c (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)butyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0253] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.b.iii (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5c was obtained in a yield of 20% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0254] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 7.98-7.88 (m, 1H), 7.56-7.51 (m, 1H), 7.50-7.44 (m, 2H), 7.42-7.18 (m, 8H), 7.13-7.04 (m, 1H), 6.98-6.90 (m, 2H), 6.85-6.78 (m, 1H), 5.05-4.99 (m, 1H), 4.81-4.72 (m, 2H), 4.71-4.64 (m, 1H), 4.25 (d, J = 15.5 Hz, 1H), 3.97-3.89 (m, 5H), 3.89-3.76 (m, 1H), 3.69-3.60 (m, 1H), 3.41-3.35 (m, 1H), 3.27-3.20 (m, 2H), 3.18-3.09 (m, 2H), 3.10-3.02 (m, 2H), 2.91-2.82 (m, 1H), 2.60-2.50 (m, 2H), 2.48-2.31 (m, 1H), 2.26-2.18 (m, 1H), 2.13-2.02 (m, 1H), 2.01-1.95 (m, 1H), 1.95-1.64 (m, 3H), 1.59-1.41 (m, 4H), 1.18-1.13 (m, 6H), 1.02 (d, J = 16.6 Hz, 9H).

[0255] 4-5. 화합물 5d (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0256] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.b.iv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5d was obtained in a yield of 20% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0257] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.14 (d, J = 9.0 Hz, 1H), 7.94-7.85 (m, 1H), 7.56-7.51 (m, 1H), 7.49-7.44 (m, 2H), 7.41-7.19 (m, 8H), 7.10-7.04 (m, 1H), 6.97-6.89 (m, 2H), 6.84-6.79 (m, 1H), 5.04-4.99 (m, 1H), 4.80-4.73 (m, 2H), 4.70-4.64 (m, 1H), 4.26 (d, J = 15.4 Hz, 1H), 3.97-3.89 (m, 5H), 3.88-3.75 (m, 1H), 3.68-3.61 (m, 1H), 3.41-3.35 (m, 1H), 3.27-3.21 (m, 2H), 3.14-3.08 (m, 2H), 3.08-2.97 (m, 2H), 2.90-2.82 (m, 1H), 2.59-2.50 (m, 2H), 2.48-2.39 (m, 1H), 2.26-2.18 (m, 1H), 2.12-2.03 (m, 1H), 2.01-1.95 (m, 1H), 1.94-1.64 (m, 3H), 1.59-1.49 (m, 2H), 1.45-1.37 (m, 2H), 1.37-1.28 (m, 2H), 1.18-1.12 (m, 6H), 1.01 (d, J = 17.1 Hz, 9H).

[0258] 4-6. 화합물 5e (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0259] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.bv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5e was obtained in a yield of 24% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0260] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.15 (d, J = 9.1, 3.5 Hz, 1H), 7.93-7.83 (m, 1H), 7.56-7.52 (m, 1H), 7.50-7.45 (m, 2H), 7.42-7.19 (m, 8H), 7.11-7.04 (m, 1H), 6.98-6.89 (m, 2H), 6.85-6.78 (m, 1H), 5.05-4.99 (m, 1H), 4.82-4.72 (m, 2H), 4.71-4.64 (m, 1H), 4.26 (d, J = 15.5 Hz, 1H), 3.97-3.89 (m, 5H), 3.88-3.76 (m, 1H), 3.68-3.61 (m, 1H), 3.41-3.35 (m, 1H), 3.28-3.20 (m, 2H), 3.16-3.07 (m, 2H), 3.06-2.96 (m, 2H), 2.91-2.82 (m, 1H), 2.60-2.50 (m, 2H), 2.48-2.39 (m, 1H), 2.26-2.18 (m, 1H), 2.12-2.03 (m, 1H), 2.02-1.95 (m, 1H), 1.95-1.64 (m, 3H), 1.57-1.49 (m, 2H), 1.41-1.21 (m, 6H), 1.18-1.11 (m, 6H), 1.02 (d, J = 17.5 Hz, 9H).

[0261] 4-7. 화합물 5f (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0262] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.ci (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5f was obtained in a yield of 26% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0263] 1H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.21-8.11 (m, 2H), 7.57-7.49 (m, 1H), 7.49-7.44 (m, 2H), 7.41-7.19 (m, 8H), 7.15-7.09 (m, 1H), 7.02-6.91 (m, 2H), 6.75-6.69 (m, 1H), 5.06-5.00 (m, 1H), 4.80-4.71 (m, 2H), 4.70-4.65 (m, 1H), 4.26-4.20 (m, 1H), 3.98-3.87 (m, 5H), 3.87-3.74 (m, 1H), 3.68-3.59 (m, 1H), 3.43-3.37 (m, 1H), 3.37-3.34 (m, 2H), 3.28-3.17 (m, 4H), 2.90-2.82 (m, 1H), 2.60-2.50 (m, 2H), 2.42-2.32 (m, 1H), 2.27-2.16 (m, 1H), 2.09-2.01 (m, 1H), 2.01-1.95 (m, 1H), 1.95-1.63 (m, 3H), 1.18-1.12 (m, 6H), 1.02 (d, J = 15.2 Hz, 9H).

[0264] 4-8. 화합물 5g (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0265] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.c.ii (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then 5g of the compound was obtained in a yield of 22% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0266] 1H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 8.04-7.95 (m, 1H), 7.58-7.50 (m, 1H), 7.49-7.44 (m, 2H), 7.42-7.19 (m, 8H), 7.08-7.03 (m, 1H), 7.02-6.92 (m, 2H), 6.69-6.61 (m, 1H), 5.07-5.01 (m, 1H), 4.82-4.73 (m, 2H), 4.70-4.65 (m, 1H), 4.29-4.22 (m, 1H), 3.98-3.89 (m, 5H), 3.88-3.78 (m, 1H), 3.69-3.60 (m, 1H), 3.43-3.36 (m, 1H), 3.32-3.19 (m, 4H), 3.15-3.05 (m, 2H), 2.91-2.82 (m, 1H), 2.61-2.50 (m, 2H), 2.48-2.33 (m, 1H), 2.29-2.19 (m, 1H), 2.12-2.04 (m, 1H), 2.04-1.98 (m, 1H), 1.98-1.72 (m, 3H), 1.71-1.61 (m, 2H), 1.15 (t, J = 7.0 Hz, 6H), 1.02 (d, J = 18.0 Hz, 9H).

[0267] 4-9. 화합물 5h (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0268] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.c.iii (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5h was obtained in a yield of 27% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0269] 1H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.17-8.12 (m, 1H), 7.97-7.89 (m, 1H), 7.56-7.50 (m, 1H), 7.49-7.44 (m, 2H), 7.42-7.19 (m, 8H), 7.09-7.04 (m, 1H), 7.01-6.92 (m, 2H), 6.56-6.50 (m, 1H), 5.07-5.01 (m, 1H), 4.80-4.73 (m, 2H), 4.71-4.65 (m, 1H), 4.24 (d, J = 15.5 Hz, 1H), 3.98-3.88 (m, 5H), 3.88-3.76 (m, 1H), 3.68-3.60 (m, 1H), 3.41-3.35 (m, 1H), 3.31-3.20 (m, 4H), 3.12-3.00 (m, 2H), 2.91-2.82 (m, 1H), 2.61-2.50 (m, 2H), 2.48-2.32 (m, 2H), 2.25-2.18 (m, 1H), 2.12-1.96 (m, 2H), 1.96-1.84 (m, 1H), 1.83-1.64 (m, 1H), 1.58-1.49 (m, 2H), 1.48-1.40 (m, 2H), 1.15 (t, J = 7.0, 1.0 Hz, 6H), 1.02 (d, J = 18.0 Hz, 9H).

[0270] 4-10. 화합물 5i (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0271] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.c.iv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5i was obtained in a yield of 24% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0272] 1H NMR (600 MHz, DMSO) δ 11.10 (s, 1H), 8.19-8.14 (m, 1H), 7.96-7.87 (m, 1H), 7.59-7.53 (m, 1H), 7.51-7.46 (m, 2H), 7.43-7.21 (m, 8H), 7.10-7.05 (m, 1H), 7.03-6.99 (m, 1H), 6.99-6.93 (m, 1H), 6.55-6.49 (m, 1H), 5.08-5.02 (m, 1H), 4.82-4.75 (m, 2H), 4.72-4.66 (m, 1H), 4.27 (d, J = 15.4 Hz, 1H), 3.99-3.90 (m, 5H), 3.90-3.78 (m, 1H), 3.69-3.62 (m, 1H), 3.42-3.38 (m, 1H), 3.31-3.21 (m, 4H), 3.10-2.98 (m, 2H), 2.92-2.83 (m, 1H), 2.62-2.52 (m, 2H), 2.49-2.32 (m, 2H), 2.27-2.20 (m, 1H), 2.14-1.99 (m, 2H), 1.99-1.86 (m, 1H), 1.85-1.66 (m, 1H), 1.60-1.52 (m, 2H), 1.46-1.39 (m, 2H), 1.36-1.28 (m, 2H), 1.17 (t, J = 7.0, 1.2 Hz, 6H), 1.03 (d, J = 17.7 Hz, 9H).

[0273] 4-11. 화합물 5j (diethyl (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonate) 제조

[0274] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1-2 hours. Then, the reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 4 (1 eq), the amine crude Int. 2.cv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. Then, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then compound 5j was obtained in a yield of 15% by reverse-phase silica column chromatography using an H2O / MeOH solvent.

[0275] 1H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.17-8.12 (m, 1H), 7.92-7.83 (m, 1H), 7.58-7.49 (m, 1H), 7.49-7.44 (m, 2H), 7.41-7.18 (m, 8H), 7.08-7.04 (m, 1H), 7.02-6.97 (m, 1H), 6.97-6.92 (m, 1H), 6.54-6.48 (m, 1H), 5.07-5.00 (m, 1H), 4.81-4.72 (m, 2H), 4.70-4.65 (m, 1H), 4.25 (d, J = 15.5 Hz, 1H), 3.98-3.88 (m, 5H), 3.87-3.75 (m, 1H), 3.68-3.61 (m, 1H), 3.40-3.35 (m, 1H), 3.29-3.19 (m, 4H), 3.08-2.94 (m, 2H), 2.91-2.82 (m, 1H), 2.61-2.50 (m, 2H), 2.48-2.38 (m, 1H), 2.36 (s, 1H), 2.26-2.17 (m, 1H), 2.13-1.98 (m, 2H), 1.97-1.85 (m, 1H), 1.84-1.64 (m, 1H), 1.58-1.49 (m, 2H), 1.40-1.33 (m, 2H), 1.34-1.23 (m, 4H), 1.18-1.13 (t, J = 7.0, 1.2 Hz, 6H), 1.01 (d, J = 17.7 Hz, 9H).

[0276] <제조예 5> PROTAC 기반 화합물 제조 (1)

[0277] 5-1. Compound SD-2737 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)amino)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxo prop-1-en-1-yl)benzyl)phosphonic acid) manufacturing

[0278] The DCM solution of compound 5a (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2737 in a yield of approximately 24%.

[0279] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.13 (d, J = 9.0 Hz, 1H), 8.11-8.01 (m, 1H), 7.57-7.52 (m, 1H), 7.48-7.41 (m, 2H), 7.41-7.36 (m, 2H), 7.35-7.31 (m, 1H), 7.29-7.19 (m, 6H), 6.98-6.90 (m, 2H), 6.87-6.82 (m, 1H), 5.04-4.99 (m, 1H), 4.79-4.73 (m, 2H), 4.69-4.64 (m, 1H), 4.24 (d, J = 15.4 Hz, 1H), 3.94-3.89 (m, 1H), 3.82-3.75 (m, 1H), 3.68-3.60 (m, 1H), 3.44-3.37 (m, 1H), 3.24-3.17 (m, 4H), 3.01-2.93 (m, 2H), 2.90-2.81 (m, 1H), 2.59-2.50 (m, 2H), 2.44-2.31 (m, 1H), 2.28-2.18 (m, 1H), 2.11-2.02 (m, 1H), 2.01-1.95 (m, 1H), 1.94-1.84 (m, 1H), 1.83-1.75 (m, 1H), 1.71-1.64 (m, 1H), 1.02 (d, J = 15.5 Hz, 9H).

[0280] 5-2. 화합물 SD-2739 ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)amino)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) amino)-3-oxo prop-1-en-1-yl)benzyl)phosphonic acid) 제조

[0281] The DCM solution of compound 5b (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2739 in a yield of approximately 21%.

[0282] 1 H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.13 (d, J = 9.0 Hz, 1H), 8.02-7.93 (m, 1H), 7.56-7.52 (m, 1H), 7.46-7.41 (m, 2H), 7.41-7.35 (m, 2H), 7.35-7.31 (m, 1H), 7.29-7.19 (m, 6H), 6.96-6.90 (m, 2H), 6.85-6.79 (m, 1H), 5.04-4.99 (m, 1H), 4.80-4.73 (m, 2H), 4.69-4.63 (m, 1H), 4.24 (d, J = 15.5 Hz, 1H), 3.94-3.89 (m, 1H), 3.88-3.77 (m, 1H), 3.69-3.60 (m, 1H), 3.41-3.34 (m, 1H), 3.18-3.10 (m, 4H), 3.00-2.93 (m, 2H), 2.90-2.82 (m, 1H), 2.59-2.54 (m, 1H), 2.52-2.50 (m, 1 H), 2.45-2.33 (m, 1H), 2.27-2.19 (m, 1H), 2.10-2.03 (m, 1H), 2.00-1.95 (m, 1H), 1.94-1.83 (m, 1H), 1.83-1.74 (m, 1H), 1.72-1.61 (m, 3H), 1.01 (d, J = 16.4 Hz, 9H).

[0283] 5-3. Compound SD-2741 (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)amino)butyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxo prop-1-en-1-yl)benzyl)phosphonic acid) manufacturing

[0284] The DCM solution of compound 5c (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2741 in a yield of approximately 27%.

[0285] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.16-8.10 (m, 1H), 7.98-7.88 (m, 1H), 7.56-7.50 (m, 1H), 7.47-7.41 (m, 2H), 7.41-7.36 (m, 2H), 7.35-7.31 (m, 1H), 7.30-7.18 (m, 6H), 6.96-6.90 (m, 2H), 6.84-6.79 (m, 1H), 5.04-4.99 (m, 1H), 4.80-4.73 (m, 2H), 4.69-4.64 (m, 1H), 4.23 (d, J = 15.5 Hz, 1H), 3.94-3.88 (m, 1H), 3.87-3.77 (m, 1H), 3.67-3.61 (m, 1H), 3.41-3.32 (m, 1H), 3.15-3.10 (m, 2H), 3.09-3.02 (m, 2H), 3.00-2.93 (m, 2H), 2.89-2.82 (m, 1H), 2.59-2.50 (m, 2H), 2.44-2.31 (m, 1H), 2.25-2.18 (m, 1H), 2.11-2.03 (m, 1H), 2.00-1.94 (m, 1H), 1.94-1.84 (m, 1H), 1.82-1.73 (m, 1H), 1.71-1.63 (m, 1H), 1.57-1.49 (m, 2H), 1.49-1.43 (m, 2H), 1.01 (d, J = 16.4 Hz, 9H).

[0286] 5-4. 화합물 SD-2743 ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonic acid) 제조

[0287] The DCM solution of compound 5d (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2743 in a yield of approximately 21%.

[0288] 1 H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.14 (d, J = 9.1 Hz, 1H), 7.94-7.85 (m, 1H), 7.57-7.50 (m, 1H), 7.48-7.41 (m, 2H), 7.41-7.36 (m, 2H), 7.35-7.31 (m, 1H), 7.30-7.17 (m, 6H), 6.93 (m, 2H), 6.85-6.77 (m, 1H), 5.05-4.97 (m, 1H), 4.81-4.72 (m, 2H), 4.71-4.62 (m, 1H), 4.25 (d, J = 15.4 Hz, 1H), 3.95-3.88 (m, 1H), 3.88-3.75 (m, 1H), 3.69-3.60 (m, 1H), 3.41-3.33 (m, 1H), 3.14-3.07 (m, 2H), 3.07-3.00 (m, 2H), 2.99-2.92 (m, 2H), 2.90-2.81 (m, 1H), 2.59-2.54 (m, 1H), 2.52-2.49 (m, 1H), 2.46-2.40 (m, 1H), 2.26-2.17 (m, 1H), 2.12-2.02 (m, 1H), 2.01-1.94 (m, 1H), 1.94-1.83 (m, 1H), 1.83-1.73 (m, 1H), 1.71-1.63 (m, 1H), 1.59-1.49 (m, 2H), 1.45-1.37 (m, 2H), 1.37-1.28 (m, 2H), 1.01 (d, J = 17.1 Hz, 9H).

[0289] 5-5. Compound SD-2745 ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxo prop-1-en-1-yl)benzyl)phosphonic acid) manufacturing

[0290] The DCM solution of compound 5e (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2745 in a yield of approximately 26%.

[0291] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.13 (d, J = 9.1 Hz, 1H), 7.92-7.84 (m, 1H), 7.56-7.51 (m, 1H), 7.46-7.41 (m, 2H), 7.40-7.36 (m, 2H), 7.35-7.31 (m, 1H), 7.29-7.18 (m, 6H), 6.95-6.89 (m, 2H), 6.84-6.79 (m, 1H), 5.04-4.99 (m, 1H), 4.78-4.73 (m, 2H), 4.69-4.64 (m, 1H), 4.25 (d, J = 15.4 Hz, 1H), 3.94-3.88 (m, 1H), 3.87-3.76 (m, 1H), 3.68-3.61 (m, 1H), 3.39-3.32 (m, 1H), 3.15-3.08 (m, 2H), 3.07-2.92 (m, 4H), 2.90-2.82 (m, 1H), 2.59-2.50 (m, 2H), 2.45-2.39 (m, 1H), 2.25-2.17 (m, 1H), 2.11-2.02 (m, 1H), 2.01-1.95 (m, 1H), 1.94-1.83 (m, 1H), 1.83-1.75 (m, 1H), 1.70-1.64 (m, 1 H), 1.57-1.49 (m, 2H), 1.41-1.29 (m, 4H), 1.29-1.20 (m, 2H), 1.01 (d, J = 17.4 Hz, 9H).

[0292] 5-6. 화합물 SD-2738 (4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino) -3-oxo prop-1-en-1-yl)benzyl)phosphonic acid) 제조

[0293] The DCM solution of compound 5f (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2738 in a yield of approximately 29%.

[0294] 1 H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.21-8.11 (m, 2H), 7.56-7.49 (m, 1H), 7.46-7.41 (m, 2H), 7.40-7.35 (m, 2H), 7.36-7.31 (m, 1H), 7.30-7.19 (m, 6H), 7.15-7.09 (m, 1H), 7.02-6.97 (m, 1H), 6.96-6.89 (m, 1H), 5.05-4.99 (m, 1H), 4.79-4.71 (m, 2H), 4.69-4.64 (m, 1H), 4.25-4.20 (m, 1H), 3.94-3.88 (m, 1H), 3.81-3.74 (m, 1H), 3.68-3.60 (m, 1H), 3.42-3.31 (m, 3H), 3.24-3.17 (m, 2H), 3.01-2.93 (m, 2H), 2.90-2.81 (m, 1H), 2.59-2.50 (m, 2H), 2.41-2.32 (m, 1H), 2.26-2.16 (m, 1H), 2.09-2.01 (m, 1H), 2.01-1.94 (m, 1H), 1.93-1.82 (m, 1H), 1.82-1.70 (m, 1H), 1.69-1.62 (m, 1H), 1.01 (d, J = 14.8 Hz, 9H).

[0295] 5-7. Compound SD-2740 ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonic acid) manufacturing

[0296] A DCM solution of 5 g (1 eq) of the compound was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain the compound SD-2740 in a yield of approximately 27%.

[0297] 1H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.14 (d, J = 8.9 Hz, 1H), 8.03-7.95 (m, 1H), 7.58-7.50 (m, 1H), 7.48-7.41 (m, 2H), 7.41-7.36 (m, 2H), 7.35-7.31 (m, 1H), 7.31-7.19 (m, 6H), 7.09-7.03 (m, 1H), 7.02-6.97 (m, 1H), 6.95-6.89 (m, 1H), 5.06-5.00 (m, 1H), 4.81-4.74 (m, 2H), 4.69-4.64 (m, 1H), 4.25 (d, J = 15.5 Hz, 1H), 3.94-3.89 (m, 1H), 3.88-3.78 (m, 1H), 3.68-3.60 (m, 1H), 3.42-3.35 (m, 1H), 3.33-3.24 (m, 2H), 3.14-3.04 (m, 2H), 3.01-2.93 (m, 2H), 2.90-2.82 (m, 1H), 2.61-2.50 (m, 2H), 2.46-2.40 (m, 1H), 2.28-2.19 (m, 1H), 2.12-2.04 (m, 1H), 2.04-1.96 (m, 1H), 1.96-1.84 (m, 1H), 1.83-1.75 (m, 1H), 1.71-1.60 (m, 3H), 1.01 (d, J = 17.7 Hz, 9H).

[0298] 5-8. 화합물 SD-2742 ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)butyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino) -3-oxo prop-1-en-1-yl)benzyl)phosphonic acid) 제조

[0299] The DCM solution of compound 5h (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2742 in a yield of approximately 23%.

[0300] 1 H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.16-8.11 (m, 1H), 7.97-7.88 (m, 1H), 7.56-7.50 (m, 1H), 7.46-7.42 (m, 2H), 7.40-7.38 (m, 2H), 7.35-7.31 (m, 1H), 7.29-7.18 (m, 6H), 7.09-7.05 (m, 1H), 7.01-6.97 (m, 1H), 6.95-6.90 (m, 1H), 5.06-4.99 (m, 1H), 4.79-4.72 (m, 2H), 4.69-4.64 (m, 1H), 4.26-4.20 (m, 1H), 3.94-3.88 (m, 1H), 3.87-3.76 (m, 1H), 3.68-3.60 (m, 1H), 3.40-3.32 (m, 1H), 3.31-3.23 (m, 2H), 3.09-3.01 (m, 2H), 3.00-2.93 (m, 2H), 2.90-2.82 (m, 1H), 2.60-2.50 (m, 2H), 2.47-2.37 (m, 1H), 2.25-2.17 (m, 1H), 2.10-2.03 (m, 1H), 2.03-1.96 (m, 1H), 1.95-1.83 (m, 1H), 1.83-1.73 (m, 1H), 1.70-1.63 (m, 1H), 1.57-1.48 (m, 2H), 1.48-1.40 (m, 2H), 1.01 (d, J = 17.7 Hz, 9H).

[0301] 5-9. Compound SD-2744 ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonic acid) manufacturing

[0302] The DCM solution of compound 5i (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2744 in a yield of approximately 24%.

[0303] 1H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.16-8.11 (m, 1H), 7.94-7.85 (m, 1H), 7.57-7.52 (m, 1H), 7.46-7.41 (m, 2H), 7.41-7.36 (m, 2H), 7.35-7.31 (m, 1H), 7.29-7.18 (m, 6H), 7.08-7.04 (m, 1H), 7.01-6.98 (m, 1H), 6.95-6.90 (m, 1H), 5.05-5.00 (m, 1H), 4.79-4.73 (m, 2H), 4.69-4.64 (m, 1H), 4.25 (d, J = 15.4 Hz, 1H), 3.94-3.88 (m, 1H), 3.88-3.76 (m, 1H), 3.67-3.61 (m, 1H), 3.40-3.33 (m, 1H), 3.28-3.21 (m, 2H), 3.07-3.00 (m, 2H), 3.00-2.93 (m, 2H), 2.90-2.82 (m, 1H), 2.60-2.50 (m, 2H), 2.47-2.38 (m, 1H), 2.25-2.18 (m, 1H), 2.12-2.04 (m, 1H), 2.03-1.97 (m, 1H), 1.96-1.83 (m, 1H), 1.83-1.72 (m, 1H), 1.71-1.64 (m, 1H), 1.59-1.50 (m, 2H), 1.44-1.36 (m, 2H), 1.35-1.26 (m, 2H), 1.01 (d, J = 17.6 Hz, 9H).

[0304] 5-10. 화합물 SD-2746 ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxo prop-1-en-1-yl)benzyl)phosphonic acid) 제조

[0305] The DCM solution of compound 5j (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2746 in a yield of approximately 28%.

[0306] 1 H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.16-8.11 (m, 1H), 7.91-7.83 (m, 1H), 7.58-7.52 (m, 1H), 7.47-7.41 (m, 2H), 7.41-7.35 (m, 2H), 7.35-7.31 (m, 1H), 7.29-7.19 (m, 6H), 7.08-7.03 (m, 1H), 7.02-6.98 (m, 1H), 6.96-6.90 (m, 1H), 5.06-5.00 (m, 1H), 4.79-4.72 (m, 2H), 4.71-4.64 (m, 1H), 4.25 (d, J = 15.4 Hz, 1H), 3.94-3.88 (m, 1H), 3.88 -3.76 (m, 1H), 3.67-3.61 (m, 1H), 3.39-3.32 (m, 1H), 3.30-3.22 (m, 2H), 3.07-2.93 (m, 4H), 2.90-2.82 (m, 1H), 2.61-2.50 (m, 2H), 2.45-2.39 (m, 1H), 2.25-2.17 (m, 1H), 2.12-2.04 (m, 1H), 2.04-1.98 (m, 1H), 1.96-1.83 (m, 1H), 1.83-1.72 (m, 1H), 1.71-1.63 (m, 1H), 1.58-1.49 (m, 2H), 1.40-1.33 (m, 2H), 1.33-1.22 (m, 4H), 1.01 (d, J = 17.8 Hz, 9H).

[0307] <Preparation Example 6> Preparation of Intermediate 3 (Int. 3)

[0308] Intermediate 3 of the present invention was prepared using the following reaction formula 6.

[0309] [Reaction Equation 6]

[0310]

[0311] 6-1. Preparation of compound di (tert-butyl (3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carba moyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropyl)carbamate)

[0312] Compound d (1 eq), 3-((tert-butoxycarbonyl)amino)propanoic acid (1 eq.), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a solvent of MeOH:MC=1:15 to obtain compound di in a yield of 79%.

[0313] 1H NMR (600 MHz, DMSO) δ 8.97 (s, 1H), 8.56 (t, J = 6.1 Hz, 1H), 7.92 (d, J = 9.3 Hz, 1H), 7.41 (d, J = 8.4 Hz, 2H), 7.37 (d, J = 8.3 Hz, 2H), 6.67 (t, J = 5.6 Hz, 1H), 5.12 (d, J = 3.4 Hz, 1H), 4.51 (d, J = 9.3 Hz, 1H), 4.44-4.39 (m, 2H), 4.35-4.31 (m, 1H), 4.22-4.17 (m, 1H), 3.68-3.60 (m, 2H), 3.15-3.06 (m, 2H), 2.43 (s, 3H), 2.42-2.35 (m, 1H), 2.32-2.25 (m, 1H), 2.05-1.99 (m, 1H), 1.91-1.86 (m, 1H), 1.35 (s, 9H), 0.92 (s, 9H).

[0314] 6-2. Preparation of compound d.ii (tert-butyl (4-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carba moyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-4-oxobutyl)carbamate)

[0315] Compound d (1 eq), 4-((tert-butoxycarbonyl)amino)butanoic acid (1 eq.), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent to obtain compound d.ii in a yield of 80%.

[0316] 1H NMR (600 MHz, DMSO) δ 8.97 (s, 1H), 8.56 (t, J = 6.1 Hz, 1H), 7.88 (d, J = 9.3 Hz, 1H), 7.41 (d, J = 8.5 Hz, 2H), 7.37 (d, J = 8.4 Hz, 2H), 6.79 (t, J = 5.6 Hz, 1H), 5.11 (d, J = 3.2 Hz, 1H), 4.52 (d, J = 9.4 Hz, 1H), 4.45-4.39 (m, 2H), 4.35-4.31 (m, 1H), 4.22-4.18 (m, 1H), 3.68-3.60 (m, 2H), 2.94-2.83 (m, 2H), 2.43 (s, 3H), 2.25-2.19 (m, 1H), 2.12-2.06 (m, 1H), 2.04-1.99 (m, 1H), 1.91-1.86 (m, 1H), 1.63-1.50 (m, 2H), 1.36 (s, 9H), 0.92 (s, 9H).

[0317] 6-3. Preparation of compound d.iii (tert-butyl (5-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carba moyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-5-oxopentyl)carbamate)

[0318] Compound d (1 eq), 5-((tert-butoxycarbonyl)amino)pentanoic acid (1 eq.), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a solvent of MeOH:MC=1:15 to obtain compound d.iii in a yield of 78%.

[0319] 1H NMR (600 MHz, DMSO) δ 8.97 (s, 1H), 8.55 (t, J = 6.1 Hz, 1H), 7.82 (d, J = 9.4 Hz, 1H), 7.41 (d, J = 8.5 Hz, 2H), 7.37 (d, J = 8.4 Hz, 2H), 6.76 (t, J = 5.7 Hz, 1H), 5.11 (d, J = 3.6 Hz, 1H), 4.52 (d, J = 9.4 Hz, 1H), 4.45-4.39 (m, 2H), 4.35-4.31 (m, 1H), 4.23-4.17 (m, 1H), 3.68-3.60 (m, 2H), 2.90-2.84 (m, 2H), 2.43 (s, 3H), 2.26-2.20 (m, 1H), 2.12-2.06 (m, 1H), 2.05-1.98 (m, 1H), 1.92-1.85 (m, 1H), 1.50-1.37 (m, 2H), 1.35 (s, 9H), 1.35-1.28 (m, 2H), 0.92 (s, 9H).

[0320] 6-4. Preparation of compound d.iv (tert-butyl (6-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carba moyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-6-oxohexyl)carbamate)

[0321] Compound d (1 eq), 6-((tert-butoxycarbonyl)amino)hexanoic acid (1 eq.), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent to obtain compound d.iv in a yield of 82%.

[0322] 1 H NMR (600 MHz, DMSO) δ 8.96 (s, 1H), 8.54 (t, J = 6.1 Hz, 1H), 7.81 (d, J = 9.4 Hz, 1H), 7.40 (d, J = 8.5 Hz, 2H), 7.36 (d, J = 8.4 Hz, 2H), 6.76 (t, J = 5.7 Hz, 1H), 5.11 (d, J = 3.6 Hz, 1H), 4.50 (d, J = 9.4 Hz, 1H), 4.43-4.43 (m, 2H), 4.35-4.31 (m, 1H), 4.23-4.17 (m, 1H), 3.68-3.60 (m, 2H), 2.90-2.84 (m, 2H), 2.43 (s, 3H), 2.26-2.20 (m, 1H), 2.12-2.06 (m, 1H), 2.05-1.98 (m, 1H), 1.92-1.85 (m, 1H), 1.51-1.32 (m, 4H), 1.35 (s, 9H), 1.35-1.28 (m, 2H), 0.92 (s, 9H).

[0323] 6-5. Preparation of compound dv (tert-butyl (7-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carba moyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-7-oxoheptyl)carbamate)

[0324] Compound d (1 eq), 7-((tert-butoxycarbonyl)amino)heptanoic acid (1 eq.), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a solvent of MeOH:MC=1:15 to obtain compound dv in a yield of 68%.

[0325] 1 H NMR (600 MHz, DMSO) δ 8.97 (s, 1H), 8.55 (t, J = 6.1 Hz, 1H), 7.83 (d, J = 9.4 Hz, 1H), 7.41 (d, J = 8.5 Hz, 2H), 7.37 (d, J = 8.4 Hz, 2H), 6.74 (t, J = 5.8 Hz, 1H), 5.11 (d, J = 3.6 Hz, 1H), 4.53 (d, J = 9.4 Hz, 1H), 4.45-4.39 (m, 2H), 4.35-4.31 (m, 1H), 4.23-4.17 (m, 1H), 3.68-3.60 (m, 2H), 2.89-2.83 (m, 2H), 2.43 (s, 3H), 2.28-2.18 (m, 1H), 2.12-2.05 (m, 1H), 2.05-1.97 (m, 1H), 1.92-1.85 (m, 1H), 1.52-1.39 (m, 2H), 1.35 (s, 9H), 1.34 -1.29 (m, 2H), 1.24-1.18 (m, 4H), 0.92 (s, 9H).

[0326] 6-6. 화합물 d.vi (tert-butyl (9-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carba moyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-9-oxononyl)carbamate) 제조

[0327] Compound d (1 eq), 9-((tert-butoxycarbonyl)amino)nonanoic acid (1 eq.), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent to obtain compound d.vi in ​​a yield of 68%.

[0328] 1 H NMR (600 MHz, DMSO) δ 8.97 (s, 1H), 8.55 (t, J = 6.1 Hz, 1H), 7.83 (d, J = 9.4 Hz, 1H), 7.41 (d, J = 8.5 Hz, 2H), 7.37 (d, J = 8.4 Hz, 2H), 6.73 (t, J = 5.8 Hz, 1H), 5.11 (d, J = 3.6 Hz, 1H), 4.53 (d, J = 9.4 Hz, 1H), 4.45-4.39 (m,2H), 4.35-4.32 (m, 1H), 4.23-4.17 (m, 1H), 3.68-3.60 (m, 2H), 2.90-2.83 (m, 2H), 2.43 (s, 3H), 2.26-2.20 (m, 1H), 2.12-2.05 (m, 1H), 2.04-1.99 (m, 1H), 1.93-1.85 (m, 1H), 1.52-1.45 (m, 2H), 1.35 (s, 9H), 1.33-1.30 (m, 2H), 1.26-1.16 (m, 8H), 0.92 (s, 9H).

[0329] 6-7. Preparation of compound d.vii (tert-butyl (11-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methylthiazol-5-yl)benzyl)carba moyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-11-oxoundecyl)carbamate)

[0330] Compound d (1 eq), 11-((tert-butoxycarbonyl)amino)undecanosic acid (1 eq.), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent to obtain compound d.vii in a yield of 59%.

[0331] 1H NMR (600 MHz, DMSO) δ 8.97 (s, 1H), 8.55 (t, J = 6.1 Hz, 1H), 7.83 (d, J = 9.3 Hz, 1H), 7.41 (d, J = 8.5 Hz, 2H), 7.37 (d, J = 8.4 Hz, 2H), 6.73 (t, J = 5.8 Hz, 1H), 5.11 (d, J = 3.5 Hz, 1H), 4.53 (d, J = 9.4 Hz, 1H), 4.45-4.39 (m, 2H), 4.35-4.31 (m, 1H), 4.23-4.17 (m, 1H), 3.68-3.60 (m, 2H), 2.89-2.83 (m, 2H), 2.43 (s, 3H), 2.25-2.21 (m, 1H), 2.12-2.04 (m, 1H), 2.04-1.98 (m, 1H), 1.93-1.85 (m, 1H), 1.53-1.45 (m, 2H), 1.35 (s, 9H), 1.34-1.28 (m, 2H), 1.25-1.17 (m, 12H), 0.92 (s, 9H).

[0332] 6-8. Preparation of Intermediate Int. 3

[0333] Intermediates Int. 3.i, Int. 3.ii, Int. 3.iii, Int. 3.iv, Int. 3.v, Int. 3.vi, and Int. 3.vii were produced by treating compounds di, d.ii, d.iii, d.iv, dv, d.vi, and d.vii, respectively, with a 25% TFA / DCM solution and removing volatile components under vacuum, and the resulting amine compounds were used directly as crude materials in the next reaction step.

[0334] <Preparation Example 7> Preparation of PROTAC-based compound (2)

[0335] The PROTAC-based compounds SD-2764 to 2770 of the present invention were prepared using the following reaction scheme 7.

[0336] [Reaction Equation 7]

[0337]

[0338] 7-1. Preparation of Compound 6

[0339] LiOH-H2O (2 eq) was added to a solution of Compound 4 (1 eq) in THF / H2O (1:1) solvent, and the reaction mixture was stirred at room temperature for 1-2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Then, each amine crude Int. 3.i, Int. 3.ii, Int. 3.iii, Int. 3.iv, Int. 3.vi, or Int. 3.vii (1 eq) was mixed with the acid crude of Compound 4 (1 eq), HATU (1.5 eq), and DIPEA (3 eq), respectively, and dissolved in DMF. The mixture was stirred at room temperature overnight. After the reaction was complete, the mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then purified by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain compounds 6a, 6b, 6c, 6d, 6e, 6f, or 6g, respectively, which were used immediately in the following reaction.

[0340] 7-2. Compound SD-2764 ((4-((E)-3-(((S)-1-((S)-2-(benzyl(3-((3-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methyl thiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxopropyl)amino)- 3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl) phosphonic acid) manufacturing

[0341] The DCM solution of compound 6a (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2764 in a yield of approximately 24%.

[0342] 1 H NMR (400 MHz, DMSO) δ 8.94 (s, 1H), 8.54 (t, J = 6.2 Hz, 1H), 8.10 (d, J = 8.8 Hz, 1H), 7.97-7.81 (m, 2H), 7.38-7.28 (m, 9H), 7.27-7.14 (m, 5H), 6.90-6.85 (m, 1H), 4.79-4.68 (m, 2H), 4.66-4.62 (m, 1H), 4.50-4.46 (m, 1H), 4.42-4.35 (m, 3H), 4.30-4.28 (m, 1H), 4.23-4.13 (m, 2H), 3.90-3.85 (m, 1H), 3.83-3.73 (m, 2H), 3.64-3.55 (m, 4H), 3.52-3.45 (m, 1H), 3.36-3.27 (m, 1H), 3.19-3.13 (m, 2H), 2.88-2.81 (m, 1H), 2.40 (s, 3H), 2.38-2.30 (m, 1H), 2.29-2.23 (m, 1H), 2.22-2.14 (m, 1H), 2.10-1.92 (m, 2H), 1.91-1.81 (m, 2H), 1.79-1.73 (m, 1H), 0.98 (d, J = 11.2 Hz, 9H), 0.87 (d, J = 9.1 Hz, 9H).

[0343] 7-3. Compound SD-2765 ((4-((E)-3-(((S)-1-((S)-2-(benzyl(3-((4-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methyl thiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-4-oxobutyl)amino)-3-oxo propyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonic acid) production

[0344] The DCM solution of compound 6b (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2765 in a yield of approximately 21%.

[0345] 1H NMR (400 MHz, DMSO) δ 8.95 (s, 1H), 8.53 (t, J = 6.1 Hz, 1H), 8.11 (d, J = 9.1 Hz, 1H), 7.94-7.81 (m, 2H), 7.40-7.28 (m, 9H), 7.27-7.15 (m, 5H), 6.91-6.86 (m, 1H), 4.77-4.71 (m, 2H), 4.65 (t, J = 8.4 Hz, 1H), 4.49 (t, J = 9.0 Hz, 1H), 4.41-4.36 (m, 3H), 4.31-4.29 (m, 1H), 4.23-4.15 (m, 2H), 3.90-3.85 (m, 1H), 3.82-3.73 (m, 2H), 3.64-3.57 (m, 4H), 3.50-3.47 (m, 1H), 3.38-3.30 (m, 1H), 2.98-2.93 (m, 2H), 2.88-2.85 (m, 1H), 2.40 (s, 3H), 2.23-2.15 (m, 2H), 2.13-1.93 (m, 3H), 1.88-1.81 (m, 2H), 1.79-1.73 (m, 1H), 1.50-1.60 (m, 2H), 0.98 (d, J = 11.7 Hz, 9H), 0.89 (d, J = 8.0 Hz, 9H).

[0346] 7-4. 화합물 SD-2766 ((4-((E)-3-(((S)-1-((S)-2-(benzyl(3-((5-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methyl thiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-5-oxopentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl) phosphonic acid) 제조

[0347] The DCM solution of compound 6c (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2766 in a yield of approximately 27%.

[0348] 1 H NMR (400 MHz, DMSO) δ 8.95 (s, 1H), 8.54 (t, J = 6.0 Hz, 1H), 8.11 (d, J = 8.9 Hz, 1H), 7.91-7.78 (m, 2H), 7.39-7.28 (m, 9H), 7.26-7.15 (m, 5H), 6.91-6.85 (m, 1H), 4.76-4.71 (m, 2H), 4.67-4.63 (t, J = 8.1 Hz, 1H), 4.51-4.47 (m, 1H), 4.42-4.35 (m, 3H), 4.31-4.28 (m, 1H), 4.23-4.15 (m, 2H), 3.90-3.85 (m, 1H), 3.80-3.73 (m, 2H), 3.62-3.58 (m, 4H), 3.53-3.46 (m,1H), 3.35-3.31 (m, 1H), 3.01-2.92 (m, 2H), 2.85-2.83 (m, 1H), 2.40 (s, 3H), 2.23-2.13 (m, 2H), 2.09-1.96 (m, 3H), 1.89-1.81 (m, 2H), 1.79-1.73 (m, 1H), 1.45-1.38 (m, 2H), 1.33-1.27 (m, 2H), 0.98 (d, J = 12.3 Hz, 9H), 0.88 (d, J = 6.3 Hz, 9H).

[0349] 7-5. Compound SD-2767 ((4-((E)-3-(((S)-1-((S)-2-(benzyl(3-((6-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methyl thiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-6-oxohexyl)amino)-3-oxo propyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonic acid) production

[0350] The DCM solution of compound 6d (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2767 in a yield of approximately 21%.

[0351] 1H NMR (400 MHz, DMSO) δ 8.94 (s, 1H), 8.60-8.51 (m, 1H), 8.11 (d, J = 8.9 Hz, 1H), 7.90-7.77 (m, 2H), 7.39-7.27 (m, 9H), 7.25-7.10 (m, 5H), 6.89-6.84 (m, 1H), 4.80-4.68 (m, 2H), 4.67-4.60 (m, 1H), 4.51-4.47 (m, 1H), 4.42-4.36 (m, 2H), 4.33-4.27 (m, 1H), 4.20-4.14 (m, 2H), 3.90-3.84 (m, 1H), 3.80-3.73 (m, 2H), 3.65-3.55 (m, 4H), 3.49-3.44 (m,1H), 3.35- 3.24 (m, 1H), 2.97- 2.92 (m, 3H), 2.84-2.65 (m, 1H), 2.40 (s, 3H), 2.23-2.13 (m, 2H), 2.12-1.96 (m, 3H), 1.91-1.80 (m, 2H), 1.79-1.69 (m, 1H), 1.50-1.37 (m, 2H), 1.37-1.26 (m, 2H), 1.23-1.14 (m, 2H), 0.98 (d, J = 11.8 Hz, 9H), 0.88 (d, J = 5.0 Hz, 9H).

[0352] 7-6. 화합물 SD-2768 ((4-((E)-3-(((S)-1-((S)-2-(benzyl(3-((7-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methyl thiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-7-oxoheptyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl) phosphonic acid) 제조

[0353] The DCM solution of compound 6e (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2768 in a yield of approximately 26%.

[0354] 1 H NMR (400 MHz, DMSO) δ 8.95 (s, 1H), 8.54 (m, 1H), 8.10 (d, J = 8.8 Hz, 1H), 7.89-7.76 (m, 2H), 7.46-7.27 (m, 9H), 7.27-7.13 (m, 5H), 6.92-6.83 (m, 1H), 4.78-4.68 (m, 2H), 4.64 (t, J = 8.0 Hz, 1H), 4.51-4.48 (m, 1H), 4.42-4.35 (m, 2H), 4.34-4.26 (m, 1H), 4.24-4.13 (m, 2H), 3.91-3.85 (m, 1H), 3.81-3.72 (m, 2H), 3.64-3.58 (m, 4H), 3.52-3.45 (m, 1H), 3.36-3.30 (m, 1H) 2.99-2.90 (m, 3H), 2.86-2.74 (m, 1H), 2.40 (s, 3H), 2.24-2.13 (m, 2H), 2.10-1.94 (m, 3H), 1.90-1.79 (m, 2H), 1.77-1.69 (m, 1H)1.47-1.36 (m, 2H), 1.34-1.25 (m, 2H), 1.24-1.12 (m, 4H), 0.98 (d, J = 12.3 Hz, 9H), 0.88 (d, J = 3.9 Hz, 9H).

[0355] 7-7. Compound SD-2769 ((4-((E)-3-(((S)-1-((S)-2-(benzyl(3-((9-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methyl thiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-9-oxononyl)amino)-3-oxo propyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)phosphonic acid) production

[0356] The DCM solution of compound 6f (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2769 in a yield of approximately 29%.

[0357] 1H NMR (400 MHz, DMSO) δ 8.95 (s, 1H), 8.53 (t, J = 6.0 Hz, 1H), 8.11 (d, J = 8.8 Hz, 1H), 7.89-7.77 (m, 2H), 7.43-7.27 (m, 9H), 7.26-7.15 (m, 5H), 6.91-6.85 (m, 1H), 4.78-4.69 (m, 2H), 4.64 (t, J = 8.2 Hz, 1H), 4.52-4.46 (m, 1H), 4.43-4.33 (m, 2H), 4.33-4.27 (m, 1H), 4.25-4.14 (m, 2H), 3.92-3.84 (m, 1H), 3.81-3.74 (m, 2H), 3.66-3.56 (m, 4H), 3.54-3.46 (m, 1H), 3.36-3.28 (m, 1H), 3.04-2.86 (m, 4H), 2.40 (s, 3H), 2.24-2.13 (m, 2H), 2.10-1.94 (m, 3H), 1.91-1.81 (m, 2H), 1.79-1.70 (m, 1H), 1.49-1.36 (m, 2H), 1.35-1.26 (m, 2H), 1.22-1.11 (m, 8H), 0.98 (d, J = 12.1 Hz, 9H), 0.89 (d, J = 2.2 Hz, 9H).

[0358] 7-8. 화합물 SD-2770 ((4-((E)-3-(((S)-1-((S)-2-(benzyl(3-((11-(((S)-1-((2S,4R)-4-hydroxy-2-((4-(4-methyl thiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-11-oxoundecyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl) phosphonic acid) 제조

[0359] A DCM solution of 6 g (1 eq) of compound was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified by reverse-phase silica column chromatography using H2O / MeOH solvent to obtain compound SD-2770 in a yield of approximately 27%.

[0360] 1 H NMR (400 MHz, DMSO) δ 8.66 (s, 1H), 7.88-7.68 (m, 1H), 7.56-7.48 (m, 1H), 7.40-7.27 (m, 6H), 7.27-7.16 (m, 5H), 7.15-7.05 (m, 2H), 6.94-6.80 (m, 1H), 6.75-6.63 (m, 1H), 6.60-6.46 (m, 1H), 6.45-6.32 (m, 1H), 5.06 (t, J = 12.8 Hz, 1H), 4.90-4.74 (m, 1H), 4.74-4.63 (m, 1H), 4.61-4.54 (m, 1H), 4.50-4.39 (m, 3H), 4.38-4.28 (m, 2H), 4.05-3.92 (m, 3H), 3.91-3.79 (m, 2H), 3.66-3.57 (m, 2H), 3.56-3.46 (m, 1H), 3.28-3.00 (m, 3H), 2.99-2.93 (m, 1H), 2.93-2.84 (m, 1H), 2.47 (s, 3H), 2.41-2.34 (m, 1H), 2.32-2.24 (m, 1H), 2.20-2.02 (m, 4H), 1.96-1.85 (m, 1H), 1.78-1.65 (m, 1H), 1.62-1.37 (m, 4H), 1.33-1.11 (m, 12H), 1.08 (d, J = 10.4 Hz, 9H), 0.93 (d, J = 3.9 Hz, 9H).

[0361] <Preparation Example 8> Preparation of Intermediate 4 (Int. 4) and Intermediate 5 (Int. 5)

[0362] Intermediate 4 of the present invention was prepared using the following reaction formula 8, and intermediate 5 was prepared using the following reaction formula 9.

[0363] [Reaction Equation 8]

[0364]

[0365] [Reaction Equation 9]

[0366]

[0367] 8-1. Preparation of Compound e (4-iodobenzyl)phosphonic acid

[0368] A solution of diethyl(4-iodobenzyl)phosphonate (1 eq) dissolved in DCM was cooled to 0 °C in an ice bath, after which CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, after which the solvent was removed under reduced pressure. The residue was purified by primary purification using reverse-phase silica column chromatography with an H2O / MeOH solvent system to obtain phosphonic acid compound e in a yield of approximately 79%, which was used immediately in the following reaction.

[0369] 8-2. Preparation of compound ei (((4-iodobenzyl)phosphoryl)bis(oxy))bis(methylene) bis(2,2-dimethylpropanoate)

[0370] Compound e (1 eq) was dissolved in DMF under a nitrogen atmosphere, and then POM-Cl (6 eq) and DIPEA (6 eq) were added. The reaction mixture was heated overnight at 60 °C. Afterward, ethyl acetate and water were added to the mixture, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under reduced pressure. The residue was purified using silica column chromatography with EA:HEX=1:5 solvent to obtain compound ei in approximately 73% yield.

[0371] 1H NMR (600 MHz, DMSO) δ 7.67-7.62 (m, 2H), 7.07-7.03 (m, 2H), 5.58-5.50 (m, 4H), 3.32 (s, 2H), 1.12 (s, 18H).

[0372] 8-3. Compound e.ii Preparation of (((hydroxy(4-iodobenzyl)phosphoryl)oxy)methyl pivalate)

[0373] LiOH·H2O (3 eq) was added to a solution of compound ei (1 eq) dissolved in a THF:H2O=1:1 mixed solvent, and the reaction mixture was stirred at room temperature for 1-2 hours. Afterward, the reaction mixture was concentrated to obtain an acidic compound e.ii, which was used directly in the next reaction step.

[0374] 1 H NMR (600 MHz, DMSO) δ 7.53-7.48 (m, 2H), 7.06-7.01 (m, 2H), 5.34 (d, J = 12.0 Hz, 2H), 2.65 (d, J = 20.5 Hz, 2H), 1.12 (s, 9H).

[0375] 8-4. Preparation of intermediate 4 ((E)-3-(4-((bis((pivaloyloxy)methoxy)phosphoryl)methyl)phenyl)acrylic acid)

[0376] Compound ei (1 eq), Pd(OAc)2 (5 mol%), and tri-o-tolylphosphine (5 mol%) were dissolved in DMA under a nitrogen atmosphere, after which tributylamine (3 eq) was added. Subsequently, acrylic acid (2.1 eq) was added, and the reaction mixture was heated at 110 °C for 4 hours. After the reaction was complete, ethyl acetate and water were added to extract the organic layer, which was then dried with MgSO4, filtered, and concentrated under reduced pressure. The residue was purified using silica column chromatography with a solvent composition of MeOH:DCM = 1:15 to obtain intermediate Int. 4 in a 45% yield.

[0377] 1 H NMR (600 MHz, DMSO) δ 7.64-7.58 (m, 2H), 7.54 (d, J = 16.0 Hz, 1H), 7.31-7.25 (m, 2H), 6.50 (d, J = 16.0 Hz, 1H), 5.59-5.51 (m, 4H), 3.41 (d, J = 22.6 Hz, 2H), 1.12 (s, 18H).

[0378] 8-5. Preparation of intermediate 5 ((E)-3-(4-((hydroxy((pivaloyloxy)methoxy)phosphoryl)methyl)phenyl)acrylic acid)

[0379] Compounds e.ii (1 eq), Pd(OAc)2 (5 mol%), and tri-o-tolylphosphine (5 mol%) were dissolved in DMA under a nitrogen atmosphere, after which tributylamine (3 eq) was added. Subsequently, acrylic acid (2.1 eq) was added, and the reaction mixture was heated at 110 °C for 4 hours. After the reaction was complete, ethyl acetate and water were added to extract the organic layer, which was then dried with MgSO4, filtered, and concentrated under reduced pressure. The residue was purified using silica column chromatography with a solvent composition of MeOH:DCM = 1:15 to obtain intermediate Int. 5 in a 33% yield.

[0380] 1 H NMR (600 MHz, DMSO) δ 7.53-7.48 (m, 3H), 7.28-7.23 (m, 2H), 6.44 (d, J = 15.9 Hz, 1H), 5.40 (d, J = 12.2 Hz, 2H), 2.83 (d, J = 20.9 Hz, 2H), 1.14 (s, 9H).

[0381] <Preparation Example 9> Preparation of PROTAC-based compound (3)

[0382] The PROTAC-based compounds SD-2902 and 2909 of the present invention were prepared using the following reaction scheme 10.

[0383] [Reaction Equation 10]

[0384]

[0385] 9-1. Compound 7 (tert-butyl ((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)carbamate) manufacturing

[0386] LiOH-H2O (2 eq) was added to a solution of Compound 3 (1 eq) in THF / H2O (1:1) solvent, and the resulting mixture was stirred at room temperature for 1–2 hours. The mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. The acid crude of Compound 3 (1 eq), the amine crude Int. 2.c.iv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF. The reaction mixture was stirred overnight at room temperature. After the reaction was complete, ethyl acetate and water were added to the mixture, and the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using MeOH:DCM = 1:15 solvent to obtain Compound 7 in a yield of approximately 49%.

[0387] 1H NMR (600 MHz, DMSO) δ 11.10 (s, 1H), 7.98-7.85 (m, 1H), 7.59-7.54 (m, 1H), 7.43-7.35 (m, 1H), 7.31-7.19 (m, 4H), 7.09 (d, J = 8.6 Hz, 1H), 7.02 (d, J = 7.0 Hz, 1H), 6.56-6.49 (m, 2H), 5.08-5.01 (m, 1H), 4.82-4.72 (m, 2H), 4.69 (d, J = 15.5 Hz, 1H), 4.32 (d, J = 15.5 Hz, 1H), 4.23-4.17 (m, 1H), 3.86-3.72 (m, 2H), 3.64-3.57 (m, 1H), 3.44-3.37 (m, 1H), 3.28 (q, J = 6.8 Hz, 2H), 3.08-3.00 (m, 2H), 2.92-2.84 (m, 1H), 2.62-2.52 (m, 2H), 2.49-2.42 (m, 1H), 2.28-2.19 (m, 1H), 2.12-1.98 (m, 2H), 1.97-1.84 (m, 1H), 1.84-1.73 (m, 1H), 1.61-1.52 (m, 2H), 1.46-1.40 (m, 2H), 1.37 (d, J = 11.4 Hz, 9H), 1.34-1.29 (m, 2H), 0.98 (d, J = 18.2 Hz, 9H).

[0388] 9-2. 화합물 SD-2902 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) amino)-3-oxoprop-1-en-1-yl)benzyl)phosphoryl)bis(oxy))bis(methylene) bis(2,2-dimethylpropanoate) 제조

[0389] Compound 7 was treated with 25% TFA / DCM for 1 hour at room temperature. Volatile substances were removed under vacuum, and the resulting crude compound was used directly in the next reaction step. Intermediate Int. 4 (1 eq), the amine crude compound of Compound 4 (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF and stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using a MeOH:DCM = 1:10 solvent to obtain Compound SD 2902 in a yield of 31%.

[0390] 1H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.18-8.11 (m, 1H), 7.94-7.85 (m, 1H), 7.57-7.51kk (m, 1H), 7.46 (t, J = 7.6 Hz, 2H), 7.41-7.35 (m, 2H), 7.36-7.30 (m, 1H), 7.30-7.18 (m, 5H), 7.06 (t, J = 8.7 Hz, 1H), 7.02-6.91 (m, 2H), 6.54-6.47 (m, 1H), 5.59-5.52 (m, 4H), 5.06-4.99 (m, 1H), 4.81-4.71 (m, 2H), 4.70-4.63 (m, 1H), 4.25 (d, J = 15.4 Hz, 1H), 3.94-3.87 (m, 1H), 3.87-3.75 (m, 1H), 3.68-3.60 (m, 1H), 3.57-3.49 (m, 1H), 3.42-3.35 (m, 3H), 3.28-3.20 (m, 2H), 3.08-2.95 (m, 2H), 2.91 -2.81 (m, 1H), 2.61-2.49 (m, 2H), 2.48-2.37 (m, 1H), 2.26-2.17 (m, 1H), 2.12-2.03 (m, 1H), 2.03-1.96 (m, 1H), 1.96-1.71 (m, 2H), 1.58-1.49 (m, 2H), 1.44-1.35 (m, 2H), 1.34-1.25 (m, 2H), 1.12 (d, J = 1.0 Hz, 18H), 1.01 (d, J = 17.5 Hz, 9H).

[0391] 9-3. Compound SD-2909 ((((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)benzyl)(hydroxy)phosphoryl)oxy)methyl pivalate)

[0392] Compound 7 was treated with 25% TFA / DCM for 1 hour at room temperature. Volatile substances were removed under vacuum, and the resulting crude compound was used directly in the next reaction step. Intermediate Int. 5 (1 eq), the amine crude compound of Compound 4 (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF and stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, the residue was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using a MeOH:DCM = 1:10 solvent to obtain Compound SD 2909 in a yield of 37%.

[0393] 1H NMR (600 MHz, DMSO) δ 11.11 (s, 1H), 8.14-8.07 (m, 1H), 7.97-7.87 (m, 1H), 7.58-7.51 (m, 1H), 7.40-7.30 (m, 4H), 7.28-7.18 (m, 6H), 7.06 (t, J = 8.4 Hz, 1H), 7.00 (t, J = 7.3 Hz, 1H), 6.93-6.87 (m, 1H), 6.53-6.48 (m, 1H), 5.38-5.31 (m, 2H), 5.05-4.99 (m, 1H), 4.80-4.72 (m, 2H), 4.69-4.63 (m, 1H), 4.26 (d, J = 15.5 Hz, 1H), 3.95-3.89 (m, 1H), 3.82-3.75 (m, 1H), 3.67-3.60 (m, 1H), 3.56-3.49 (m, 1H), 3.40-3.36 (m, 1H), 3.28-3.20 (m, 2H), 3.08-2.95 (m, 2H), 2.90-2.81 (m, 1H), 2.75-2.67 (m, 2H), 2.60-2.50 (m, 2H), 2.46-2.38 (m, 1H), 2.26-2.18 (m, 1H), 2.11-2.03 (m, 1H), 2.03-1.96 (m, 1H), 1.96-1.71 (m, 2H), 1.59-1.50 (m, 2H), 1.44-1.36 (m, 2H), 1.34-1.25 (m, 2H), 1.13 (d, J = 0.9 Hz, 9H), 1.01 (d, J = 17.4 Hz, 9H).

[0394] <Preparation Example 10> Preparation of Intermediate 6 (Int. 6)

[0395] Intermediate 6 of the present invention was prepared using the following reaction formula 11.

[0396] [Reaction Equation 11]

[0397]

[0398] 10-1. Preparation of compound f (ethyl (E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)acrylate)

[0399] Diethyl((4-bromophenyl)difluoromethyl)phosphonate (1 eq), Na2CO3 (2 eq), and ethyl(E)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)acrylate (1.3 eq) were dissolved in a mixed solvent of H2O / Dioxane = 1:5 by volume and stirred for about 5 minutes under a nitrogen atmosphere. Subsequently, Pd(PPh3)4 (5%) and xantphos (10%) were added, purged with nitrogen 2-3 times, and heated at 90.0 °C for 4 hours. After the reaction was complete, the mixture was mixed with ethyl acetate and water, the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex = 1:3 solvent to obtain compound f in a yield of about 54%.

[0400] 1 H NMR (600 MHz, DMSO) δ 7.89 (d, J = 8.1 Hz, 2H), 7.69 (d, J = 16.1 Hz, 1H), 7.57 (d, J = 7.0 Hz, 2H), 6.74 (d, J = 16.1 Hz, 1H), 4.19 (q, J = 7.1 Hz, 2H), 4.17-4.07 (m, 4H), 1.25 (t, J = 7.1 Hz, 3H), 1.21 (t, J = 7.0 Hz, 6H).

[0401] 10-2. Preparation of intermediate 6 ((E)-3-(4-((ethoxy(hydroxy)phosphoryl)difluoromethyl)phenyl)acrylic acid)

[0402] LiOH-H2O (2 eq) was added to a solution of compound f (1 eq) in THF / H2O (1:1) solvent, and the reaction mixture was stirred at room temperature for 1-2 hours. Afterward, the reaction mixture was concentrated, and the resulting crude residue was used directly in the next reaction step.

[0403] <Preparation Example 11> Preparation of PROTAC-based compound (4)

[0404] The PROTAC-based compounds SD-2780 to 2783 of the present invention were prepared using the following reaction scheme 12.

[0405] [Reaction Equation 12]

[0406]

[0407] 11-1. Preparation of compound 8 (ethyl 3-((2S)-N-benzyl-1-((2S)-2-((E)-3-(4-((ethoxy(hydroxy)phosphoryl) difluoro methyl)phenyl)acrylamido)-3,3-dimethylbutanoyl)pyrrolidine-2-carboxamido)propanoate)

[0408] Compound 3 was treated with 25% TFA / DCM for 1 hour at room temperature. Volatile substances were removed under vacuum, and the resulting crude product was used directly in the next reaction step. Subsequently, acid compound Int. 6 (1 eq), compound 3 (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the reaction mixture was stirred overnight at room temperature. After the reaction was complete, ethyl acetate and water were added to extract the organic layer, which was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using a MeOH:DCM = 1:15 solvent to obtain compound 8 in a yield of 73%.

[0409] 1H NMR (600 MHz, DMSO) δ 8.32-8.25 (m, 1H), 7.64 (t, J = 7.7 Hz, 2H), 7.59-7.55 (m, 2H), 7.46 (d, J = 15.7 Hz, 1H), 7.42 (d, J = 6.9 Hz, 1H), 7.36 (t, J = 7.6 Hz, 1H), 7.31-7.22 (m, 3H), 7.10-7.04 (m, 1H), 4.79-4.73 (m, 2H), 4.72-4.68 (m, 1H), 4.34 (d, J = 15.4 Hz, 1H), 4.06-3.94 (m, 4H), 3.76-3.63 (m, 2H), 3.61-3.50 (m, 2H), 2.85-2.78 (m, 1H), 2.64-2.52 (m, 1H), 2.28-2.20 (m, 1H), 2.11-2.04 (m, 1H), 2.00-1.66 (m, 3H), 1.20-1.11 (m, 6H), 1.04 (d, J = 10.4 Hz, 9H).

[0410] 11-2. 화합물 9a (ethyl hydrogen ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxo butan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonate) 제조

[0411] LiOH-H2O (2 eq) was added to a THF / H2O (1:1) solution containing compound 8 (1 eq) and stirred at room temperature for 1-2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of compound 8 (1 eq), the amine crude Int. 2.b.iv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then purified by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain compound 9a in a yield of about 40%.

[0412] 1H NMR (600 MHz, DMSO) δ 11.07 (s, 1H), 8.25-8.19 (m, 1H), 7.97-7.87 (m, 1H), 7.60-7.51 (m, 4H), 7.45-7.38 (m, 2H), 7.38-7.32 (m, 1H), 7.31-7.18 (m, 4H), 7.14-7.09 (m, 1H), 7.05-7.00 (m, 1H), 6.96-6.92 (m, 1H), 6.87-6.81 (m, 1H), 5.06-5.01 (m, 1H), 4.83-4.73 (m, 2H), 4.72-4.66 (m, 1H), 4.29 (d, J = 15.4 Hz, 1H), 3.97-3.91 (m, 1H), 3.84-3.74 (m, 2H), 3.71-3.63 (m, 1H), 3.60-3.50 (m, 1H), 3.43-3.37 (m, 1H), 3.20-3.16 (m, 1H), 3.16-3.08 (m, 3H), 3.06-2.99 (m, 2H), 2.61-2.55 (m, 1H), 2.54-2.51 (m, 1H), 2.49-2.40 (m, 1H), 2.28-2.20 (m, 1H), 2.14-2.04 (m, 1H), 2.03-1.96 (m, 1H), 1.96-1.65 (m, 3H), 1.62-1.51 (m, 2H), 1.46-1.28 (m, 4H), 1.10-1.06 (m, 3H), 1.04 (d, J = 16.8 Hz, 9H).

[0413] 11-3. 화합물 9b (ethyl hydrogen ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxo butan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonate) 제조

[0414] LiOH-H2O (2 eq) was added to a THF / H2O (1:1) solution containing Compound 8 (1 eq) and stirred at room temperature for 1–2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 8 (1 eq), the amine crude Int. 2.bv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, followed by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain Compound 9b in a yield of approximately 36%.

[0415] 1H NMR (600 MHz, DMSO) δ 11.07 (s, 1H), 8.25-8.18 (m, 1H), 7.93-7.85 (m, 1H), 7.59-7.50 (m, 4H), 7.45-7.38 (m, 2H), 7.38-7.33 (m, 1H), 7.28-7.19 (m, 4H), 7.16-7.08 (m, 1H), 7.06-7.00 (m, 1H), 6.96-6.92 (m, 1H), 6.87-6.80 (m, 1H), 5.06-5.01 (m, 1H), 4.83-4.73 (m, 2H), 4.71-4.66 (m, 1H), 4.31-4.25 (m, 1H), 3.97-3.90 (m, 1H), 3.82-3.73 (m, 2H), 3.69-3.62 (m, 1H), 3.62-3.50 (m, 1H), 3.43-3.36 (m, 1H), 3.17-3.08 (m, 3H), 3.07-2.98 (m, 3H), 2.61-2.55 (m, 1H), 2.54-2.52 (m, 1H), 2.49-2.41 (m, 1H), 2.28-2.20 (m, 1H), 2.12-2.04 (m, 1H), 2.02-1.97 (m, 1H), 1.97-1.62 (m, 3H), 1.60-1.49 (m, 2H), 1.43-1.32 (m, 4H), 1.32-1.23 (m, 2H), 1.10-1.05 (m, 3H), 1.04 (d, J = 15.5 Hz, 9H).

[0416] 11-4. 화합물 9c (ethyl hydrogen ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxo butan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonate) 제조

[0417] LiOH-H2O (2 eq) was added to a THF / H2O (1:1) solution containing compound 8 (1 eq) and stirred at room temperature for 1-2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of compound 8 (1 eq), the amine crude Int. 2.c.iv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then purified by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain compound 9c in a yield of about 34%.

[0418] 1H NMR (600 MHz, DMSO) δ 11.10 (s, 1H), 8.24-8.19 (m, 1H), 7.97-7.86 (m, 1H), 7.60-7.50 (m, 4H), 7.45-7.38 (m, 2H), 7.38-7.33 (m, 1H), 7.28-7.18 (m, 4H), 7.08 (t, J = 8.2 Hz, 1H), 7.06-6.99 (m, 2H), 6.55-6.50 (m, 1H), 5.08-5.02 (m, 1H), 4.84-4.73 (m, 2H), 4.73-4.66 (m, 1H), 4.28 (d, J = 15.5 Hz, 1H), 3.97-3.91 (m, 1H), 3.84-3.72 (m, 2H), 3.70-3.62 (m, 1H), 3.61-3.49 (m, 1H), 3.43-3.36 (m, 1H), 3.31-3.23 (m, 2H), 3.16-3.09 (m, 1H), 3.09-2.97 (m, 2H), 2.92-2.83 (m, 1H), 2.62-2.52 (m, 2H), 2.50-2.41 (m, 1H), 2.27-2.19 (m, 1H), 2.12-1.99 (m, 2H), 1.97-1.64 (m, 3H), 1.60-1.52 (m, 2H), 1.46-1.38 (m, 2H), 1.36-1.27 (m, 2H), 1.10-1.06 (m, 3H), 1.04 (d, J = 16.1 Hz, 9H).

[0419] 11-5. 화합물 9d (ethyl hydrogen ((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxo butan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonate) 제조

[0420] LiOH-H2O (2 eq) was added to a THF / H2O (1:1) solution containing Compound 8 (1 eq) and stirred at room temperature for 1–2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude of Compound 8 (1 eq), the amine crude Int. 2.cv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, followed by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain Compound 9d in a yield of approximately 39%.

[0421] 1H NMR (600 MHz, DMSO) δ 11.06 (s, 1H), 8.21-8.15 (m, 1H), 7.90-7.81 (m, 1H), 7.55-7.47 (m, 4H), 7.41-7.33 (m, 2H), 7.30 (t, J = 7.5 Hz, 1H), 7.25-7.15 (m, 4H), 7.06-6.98 (m, 2H), 6.98-6.94 (m, 1H), 6.52-6.45 (m, 1H), 5.04-4.96 (m, 1H), 4.78-4.68 (m, 2H), 4.68-4.61 (m, 1H), 4.23 (d, J = 15.6 Hz, 1H), 3.93-3.85 (m, 1H), 3.79-3.68 (m, 2H), 3.66-3.57 (m, 1H), 3.55-3.47 (m, 1H), 3.40-3.34 (m, 1H), 3.27-3.18 (m, 2H), 3.07-3.01 (m, 1H), 3.02-2.91 (m, 2H), 2.89-2.78 (m, 1H), 2.59-2.51 (m, 1H), 2.50-2.47 (m, 1H), 2.43-2.36 (m, 1H), 2.24-2.13 (m, 1H), 2.09-1.93 (m, 2H), 1.92-1.59 (m, 3H), 1.55-1.45 (m, 2H), 1.39-1.22 (m, 6H), 1.03 (t, J = 7.0 Hz, 3H), 0.99 (d, J = 11.9 Hz, 9H).

[0422] 11-6. 화합물 SD-2780 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino) -3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0423] A DCM solution containing compound 9d (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2780 in a yield of approximately 34%.

[0424] 1 H NMR (600 MHz, DMSO) δ 11.11 (s, 1H), 8.26-8.21 (m, 1H), 7.94-7.85 (m, 1H), 7.61-7.52 (m, 4H), 7.46-7.38 (m, 2H), 7.36-7.33 (m, 1H), 7.30-7.20 (m, 4H), 7.11-6.99 (m, 3H), 6.55-6.50 (m, 1H), 5.08-5.02 (m, 1H), 4.82-4.73 (m, 2H), 4.72-4.66 (m, 1H), 4.27 (d, J = 15.4 Hz, 1H), 3.96-3.91 (m, 1H), 3.84-3.77 (m, 1H), 3.69-3.63 (m, 1H), 3.58-3.51 (m, 1H), 3.41-3.34 (m, 1H), 3.31-3.24 (m, 2H), 3.11-3.07 (m, 1H), 3.07-2.96 (m, 2H), 2.63-2.52 (m, 2H), 2.47-2.41 (m, 1H), 2.27-2.19 (m, 1H), 2.14-2.06 (m, 1H), 2.06-1.99 (m, 1H), 1.98-1.65 (m, 2H), 1.60-1.51 (m, 2H), 1.41-1.36 (m, 2H), 1.36-1.25 (m, 4H), 1.03 (d, J = 17.8 Hz, 9H).

[0425] 11-7. Compound SD-2781 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino) -3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) Manufacture

[0426] A DCM solution containing compound 9b (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2781 in a yield of approximately 31%.

[0427] 1H NMR (600 MHz, DMSO) δ 11.07 (s, 1H), 8.26-8.21 (m, 1H), 7.96-7.86 (m, 1H), 7.60-7.53 (m, 4H), 7.47-7.38 (m, 2H), 7.37-7.33 (m, 1H), 7.31-7.19 (m, 4H), 7.15-7.09 (m, 1H), 7.07-7.01 (m, 1H), 6.95-6.91 (m, 1H), 6.86-6.80 (m, 1H), 5.05-5.01 (m, 1H), 4.82-4.74 (m, 2H), 4.72-4.66 (m, 1H), 4.27 (d, J = 15.4 Hz, 1H), 3.96- 3.90 (m, 1H), 3.84- 3.76 (m, 1H), 3.69-3.62 (m, 1H), 3.56- 3.51 (m, 1H), 3.41-3.36 (m, 1H), 3.16- 3.09 (m, 3H), 3.05-2.99 (m, 2H), 2.60-2.51 (m, 2H), 2.49-2.40 (m, 1H), 2.28-2.19 (m, 1H), 2.13- 2.04 (m, 1H), 2.03-1.97 (m, 1H), 1.95 -1.74 (m, 2H), 1.58-1.50 (m, 2H), 1.42-1.31 (m, 4H), 1.31-1.21 (m, 2H), 1.04 (d, J = 17.6 Hz, 9H).

[0428] 11-8. 화합물 SD-2782 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0429] A DCM solution containing compound 9c (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2782 in a yield of approximately 28%.

[0430] 1 H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.22 (d, J = 8.7 Hz, 1H), 7.95-7.83 (m, 1H), 7.58-7.43 (m, 4H), 7.42-7.33 (m, 2H), 7.32-7.26 (d, J = 7.9 Hz, 1H), 7.25-7.09 (m, 4H), 7.07-6.92 (m, 3H), 6.53-6.45 (m, 1H), 5.05-4.95 (m, 1H), 4.79-4.68 (m, 2H), 4.68-4.60 (m, 1H), 4.23 (d, J = 15.8 Hz, 1H), 3.92-3.85 (m, 1H), 3.84-3.68 (m, 1H), 3.66-3.56 (m, 1H), 3.56-3.40 (m, 1H), 3.39-3.28 (m,1H), 3.28-3.15 (m, 2H), 3.11-3.03 (m, 1H), 3.03-2.90 (m, 2H), 2.59-2.47 (m, 2H), 2.43-2.35 (m, 1H), 2.23-2.12 (m, 1H), 2.08-1.91 (m, 2H), 1.91-1.59 (m, 2H), 1.57-1.44 (m, 2H), 1.41-1.21 (m, 4H), 1.02 (d, J = 17.7 Hz, 9H).

[0431] 11-9. Compound SD-2783 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid)

[0432] A DCM solution containing compound 9a (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2783 in a yield of approximately 30%.

[0433] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.20 (d, J = 8.9 Hz, 1H), 7.96-7.86 (m, 1H), 7.55-7.43 (m, 4H), 7.42-7.33 (m, 2H), 7.32-7.27 (m, 1H), 7.26-7.14 (m, 4H), 7.13-7.07 (m, 1H), 7.02-6.93 (m, 1H), 6.91-6.84 (m, 1H), 6.82-6.74 (m, 1H), 5.03-4.95 (m, 1H), 4.79-4.68 (m, 2H), 4.68-4.60 (m, 1H), 4.23 (d, J = 15.4 Hz, 1H), 3.93-3.85 (m, 1H), 3.79-3.71 (m, 1H), 3.66-3.57 (m, 1H), 3.53-3.48 (m, 1H), 3.43-3.40 (m, 1H), 3.11-3.02 (m, 3H), 3.01-2.93 (m, 2H), 2.59-2.48 (m, 2H), 2.44-2.37 (m, 1H), 2.24-2.15 (m,1 H), 2.09-2.00 (m, 1H), 1.99-1.90 (m, 1H), 1.89-1.60 (m, 2H), 1.58-1.45 (m, 2H), 1.42-1.23 (m, 4H), 0.99 (d, J = 11.6 Hz, 9H).

[0434] <Preparation Example 12> Preparation of Intermediate 7 (Int. 7)

[0435] Intermediate 7 of the present invention was prepared using the following reaction formula 13.

[0436] [Reaction Equation 13]

[0437]

[0438] 12-1. Preparation of compound g (ethyl (E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoate)

[0439] Diethyl((4-bromophenyl)difluoromethyl)phosphonate (1 eq), Na2CO3 (2 eq), and ethyl(Z)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)but-2-enoate (1.5 eq) were dissolved in a mixed solvent of H2O:Dioxane = 1:5 by volume and stirred for about 5 minutes under a nitrogen atmosphere. Subsequently, Pd(PPh3)4 (5 mol%) and xantphos (10%) were added, and after purging with nitrogen 2-3 times, the mixture was heated at 90 °C for 4 hours. After the reaction was complete, ethyl acetate and water were mixed, and the organic layer was extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using an EA:Hex = 1:3 solvent to obtain g of the compound in a yield of about 54%.

[0440] 1 H NMR (600 MHz, DMSO) δ 7.74 (d, J = 8.3 Hz, 2H), 7.57 (d, J = 7.3 Hz, 2H), 6.22 (q, J = 1.3 Hz, 1H), 4.19-4.07 (m, 6H), 2.52 (d, J = 1.3 Hz, 3H), 1.25-1.20 (m, 9H).

[0441] 12-2. Preparation of intermediate 7 ((E)-3-(4-((diethoxyphosphoryl)difluoromethyl)phenyl)but-2-enoic acid)

[0442] LiOH-H2O (2 eq) was added to a solution of g (1 eq) of the compound in THF / H2O (1:1) solvent, and the reaction mixture was stirred at room temperature for 1-2 hours. Afterward, the reaction mixture was concentrated, and the resulting crude residue was used directly in the next reaction step.

[0443] <Preparation Example 13> Preparation of PROTAC-based compound (5)

[0444] The PROTAC-based compounds SD-2875 to 2877 of the present invention were prepared using the following reaction scheme 14.

[0445] [Reaction Equation 14]

[0446]

[0447] 13-1. Preparation of compound 10 (ethyl 3-((2S)-N-benzyl-1-((2S)-2-((E)-3-(4-((ethoxy(hydroxy)phosphoryl)difluoro methyl)phenyl)but-2-enamido)-3,3-dimethylbutanoyl)pyrrolidine-2-carboxamido)propanoate)

[0448] Compound 3 was treated with 25% TFA / DCM for 1 hour at room temperature. Volatile substances were removed under vacuum, and the resulting crude product was used directly in the next reaction step. Subsequently, acid compound Int. 7 (1 eq), compound 3 (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the reaction mixture was stirred overnight at room temperature. After the reaction was complete, ethyl acetate and water were added to extract the organic layer, which was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using a MeOH:DCM = 1:15 solvent to obtain compound 10 in a yield of 30%.

[0449] 1H NMR (600 MHz, DMSO) δ 7.58 (d, J = 8.5 Hz, 2H), 7.54 (d, J = 8.3 Hz, 2H), 7.42 (d, J = 7.2 Hz, 1H), 7.38 (t, J = 7.6 Hz, 1H), 7.34 (s, 1H), 7.33-7.28 (m, 1H), 7.28-7.22 (m, 3H), 6.18 (q, J = 1.3 Hz, 1H), 4.81-4.76 (m, 1H), 4.73-4.68 (m, 1H), 4.38 (d, J = 15.4 Hz, 1H), 4.19-4.13 (m, 2H), 4.05-3.99 (m, 2H), 3.96-3.87 (m, 1H), 3.80-3.75 (m, 2H), 3.73-3.67 (m, 1H), 3.62-3.47 (m, 1H), 2.83-2.78 (m, 1H), 2.65-2.58 (m, 1H), 2.53 (d, J = 1.3 Hz, 3H), 2.35-2.29 (m, 1H), 2.10-2.03 (m, 1H), 2.00-1.81 (m, 1H), 1.80-1.63 (m, 1H), 1.25 (t, J = 7.1 Hz, 3H), 1.16 (t, J = 7.1 Hz, 3H), 1.10-1.04 (m, 9H).

[0450] 13-2. 화합물 11a (ethyl hydrogen ((4-((E)-4-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxo butan-2-yl)amino)-4-oxobut-2-en-2-yl)phenyl)difluoromethyl)phosphonate) 제조

[0451] LiOH-H2O (2 eq) was added to a THF / H2O (1:1) solution containing compound 10 (1 eq) and stirred at room temperature for 1-2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude compound of compound 10 (1 eq), the amine crude Int. 2.b.iv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the mixture was stirred at room temperature overnight. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then purified by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain compound 11a in a yield of about 33%.

[0452] 1H NMR (600 MHz, DMSO) δ 11.03 (s, 1H), 8.14-8.09 (m, 1H), 7.99-7.89 (m, 1H), 7.57-7.51 (m, 5H), 7.40 (d, J = 6.9 Hz, 1H), 7.35 (t, J = 7.7 Hz, 1H), 7.30-7.20 (m, 3H), 7.17-7.11 (m, 1H), 6.96-6.91 (m, 1H), 6.86-6.81 (m, 1H), 6.63-6.59 (m, 1H), 5.05-5.01 (m, 1H), 4.82-4.74 (m, 2H), 4.66 (t, J = 9.1 Hz, 1H), 4.28 (d, J = 15.5 Hz, 1H), 3.98-3.92 (m, 1H), 3.85-3.74 (m, 3H), 3.70-3.62 (m, 1H), 3.58-3.51 (m, 1H), 3.44-3.39 (m, 1H), 3.16-2.97 (m, 5H), 2.61-2.52 (m, 2H), 2.49-2.41 (m, 1H), 2.29-2.20 (m, 1H), 2.14-2.04 (m, 1H), 2.03-1.96 (m, 1H), 1.96-1.75 (m, 2H), 1.61-1.51 (m, 2H), 1.46-1.39 (m, 2H), 1.39-1.29 (m, 2H), 1.08 (t, J = 7.1 Hz, 3H), 1.03 (d, J = 17.6 Hz, 9H).

[0453] 13-3. 화합물 11b (ethyl hydrogen ((4-((E)-4-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxo butan-2-yl)amino)-4-oxobut-2-en-2-yl)phenyl)difluoromethyl)phosphonate) 제조

[0454] LiOH-H2O (2 eq) was added to a THF / H2O (1:1) solution containing compound 10 (1 eq) and stirred at room temperature for 1-2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude compound of compound 10 (1 eq), the amine crude Int. 2.bv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then purified by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain compound 11b in a yield of about 32%.

[0455] 1H NMR (600 MHz, DMSO) δ 11.03 (s, 1H), 8.14-8.07 (m, 1H), 7.95-7.85 (m, 1H), 7.55-7.47 (m, 5H), 7.40-7.36 (m, 1H), 7.36-7.29 (m, 1H), 7.28-7.18 (m, 3H), 7.17-7.10 (m, 1H), 6.95-6.89 (m, 1H), 6.85-6.78 (m, 1H), 6.61-6.56 (m, 1H), 5.03-4.98 (m, 1H), 4.79-4.71 (m, 2H), 4.64 (t, J = 8.7 Hz, 1H), 4.26 (d, J = 15.5 Hz, 1H), 4.13-4.05 (m, 2H), 3.96-3.89 (m, 1H), 3.83-3.74 (m, 1H), 3.67-3.61 (m, 1H), 3.56-3.50 (m, 1H), 3.40-3.34 (m, 1H), 3.15-3.06 (m, 3H), 3.05-2.96 (m, 2H), 2.59-2.50 (m, 2H), 2.46-2.38 (m, 1H), 2.26-2.17 (m, 1H), 2.10-2.02 (m, 1H), 2.01-1.94 (m, 1H), 1.94-1.86 (m, 1H), 1.80-1.73 (m, 1H), 1.58-1.46 (m, 2H), 1.42-1.29 (m, 4H), 1.29-1.19 (m, 2H), 1.07 (t, J = 7.2 Hz, 3H), 1.01 (d, J = 18.1 Hz, 9H).

[0456] 13-4. Compound 11c (ethyl hydrogen ((4-((E)-4-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoind olin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxo butan-2-yl)amino)-4-oxobut-2-en-2-yl)phenyl)difluoromethyl)phosphonate) manufacturing

[0457] LiOH-H2O (2 eq) was added to a THF / H2O (1:1) solution containing compound 10 (1 eq) and stirred at room temperature for 1-2 hours. The reaction mixture was concentrated, and the resulting acid crude residue was used directly in the next reaction step. Subsequently, the acid crude compound of compound 10 (1 eq), the amine crude Int. 2.c.iv (1 eq), HATU (1.5 eq), and DIPEA (3 eq) were dissolved in DMF, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was then extracted, dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by normal-phase silica column chromatography using a MeOH:MC=1:15 solvent, and then purified by reverse-phase silica column chromatography using an H2O / MeOH solvent to obtain compound 11c in a yield of about 28%.

[0458] 1H NMR (600 MHz, DMSO) δ 11.04 (s, 1H), 8.12-8.06 (m, 1H), 7.97-7.86 (m, 1H), 7.55-7.46 (m, 5H), 7.37 (d, J = 7.2 Hz, 1H), 7.33 (t, J = 7.6 Hz, 1H), 7.27-7.17 (m, 3H), 7.17-7.11 (m, 1H), 6.94-6.89 (m, 1H), 6.84-6.79 (m, 1H), 6.60-6.56 (m, 1H), 5.03-4.98 (m, 1H), 4.80-4.72 (m, 2H), 4.65 (t, J = 9.3 Hz, 1H), 4.26 (d, J = 15.4 Hz, 1H), 4.13-4.05 (m, 2H) (d, J = 15.4 Hz, 1H), 3.96-3.90 (m, 1H), 3.83-3.76 (m, 1H), 3.67-3.61 (m, 1H), 3.56-3.50 (m, 1H), 3.41-3.35 (m, 1H), 3.14-3.07 (m, 2H), 3.06-2.96 (m, 3H), 2.59-2.50 (m, 2H), 2.47-2.39 (m, 1H), 2.26-2.18 (m, 1H), 2.12-2.03 (m, 1H), 2.01-1.94 (m, 1H), 1.93-1.84 (m, 1H), 1.82-1.73 (m, 1H), 1.58-1.48 (m, 2H), 1.43 -1.37 (m, 2H), 1.37-1.28 (m, 2H), 1.08 (t, J = 7.1 Hz, 3H), 1.00 (d, J = 17.7 Hz, 9H).

[0459] 13-5. Compound SD-2875 (((4-((E)-4-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-4-oxobut-2-en-2-yl)phenyl)difluoromethyl)phosphonic acid)

[0460] A DCM solution containing compound 11c (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2875 in a yield of approximately 34%.

[0461] 1H NMR (600 MHz, DMSO) δ 11.05 (s, 1H), 8.13-8.07 (m, 1H), 7.98-7.87 (m, 1H), 7.56-7.47 (m, 5H), 7.38 (d, J = 7.2 Hz, 1H), 7.33 (t, J = 7.6 Hz, 1H), 7.28-7.18 (m, 3H), 7.17-7.11 (m, 1H), 6.94-6.89 (m, 1H), 6.84-6.79 (m, 1H), 6.60-6.56 (m, 1H), 5.03-4.98 (m, 1H), 4.80-4.72 (m, 2H), 4.65 (t, J = 9.3 Hz, 1H), 4.26 (d, J = 15.4 Hz, 1H), 3.96-3.90 (m, 1H), 3.83-3.76 (m, 1H), 3.67-3.61 (m, 1H), 3.56-3.50 (m, 1H), 3.41-3.35 (m, 1H), 3.14-3.07 (m, 2H), 3.06-2.96 (m, 3H), 2.59-2.50 (m, 2H), 2.47-2.39 (m, 1H), 2.26-2.18 (m, 1H), 2.12-2.03 (m, 1H), 2.01-1.94 (m, 1H), 1.94-1.86 (m, 1H), 1.80-1.73 (m, 1H), 1.58-1.49 (m, 2H), 1.44 -1.37 (m, 2H), 1.36-1.28 (m, 2H), 1.01 (d, J = 17.7 Hz, 9H).

[0462] 13-6. 화합물 SD-2876 (((4-((E)-4-(((2S)-1-((2S)-2-(benzyl(3-((5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-5-yl)amino)pentyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl) amino)-4-oxobut-2-en-2-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0463] A DCM solution containing compound 11a (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2876 in a yield of approximately 37%.

[0464] 1 H NMR (600 MHz, DMSO) δ 11.11 (s, 1H), 8.16-8.08 (m, 1H), 7.99-7.88 (m, 1H), 7.61-7.48 (m, 5H), 7.40 (d, J = 7.2 Hz, 1H), 7.35 (t, J = 7.6 Hz, 1H), 7.30-7.20 (m, 3H), 7.11-7.06 (m, 1H), 7.04-6.99 (m, 1H), 6.62-6.57 (m, 1H), 6.56-6.50 (m, 1H), 5.07-5.02 (m, 1H), 4.82-4.74 (m, 2H), 4.67 (t, J = 9.6 Hz, 1H), 4.28 (d, J = 15.5 Hz, 1H), 3.98-3.91 (m, 1H), 3.85-3.78 (m, 1H), 3.69-3.63 (m, 1H), 3.57-3.52 (m, 1H), 3.41-3.36 (m, 1H), 3.30-3.24 (m, 2H), 3.12-3.07 (m, 1 H), 3.07-2.97 (m, 2H), 2.63-2.52 (m, 2H), 2.49-2.41 (m, 1H), 2.29-2.19 (m, 1H), 2.14-2.06 (m, 1H), 2.05-1.99 (m, 1H), 1.98-1.65 (m, 2H), 1.61-1.52 (m, 2H), 1.46-1.39 (m, 2H), 1.37-1.27 (m, 2H), 1.03 (d, J = 17.8 Hz, 9H).

[0465] 13-7. Compound SD-2877 (((4-((E)-4-(((2S)-1-((2S)-2-(benzyl(3-((6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-5-yl)amino)hexyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-4-oxobut-2-en-2-yl)phenyl)difluoromethyl)phosphonic acid)

[0466] A DCM solution containing compound 11b (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15–30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2877 in a yield of approximately 30%.

[0467] 1H NMR (600 MHz, DMSO) δ 11.03 (s, 1H), 8.13-8.06 (m, 1H), 7.94-7.85 (m, 1H), 7.56-7.47 (m, 5H), 7.40-7.36 (m, 1H), 7.36-7.29 (m, 1H), 7.28-7.18 (m, 3H), 7.17-7.10 (m, 1H), 6.95-6.89 (m, 1H), 6.85-6.78 (m, 1H), 6.61-6.56 (m, 1H), 5.03-4.98 (m, 1H), 4.79-4.71 (m, 2H), 4.64 (t, J = 8.7 Hz, 1H), 4.26 (d, J = 15.5 Hz, 1H), 3.96-3.89 (m, 1H), 3.83-3.74 (m, 1H), 3.67-3.61 (m, 1H), 3.56-3.50 (m, 1H), 3.40-3.34 (m, 1H), 3.15-3.06 (m, 3H), 3.05-2.96 (m, 2H), 2.59-2.50 (m, 2H), 2.46-2.38 (m, 1H), 2.26-2.17 (m, 1H), 2.10-2.02 (m, 1H), 2.01-1.94 (m, 1H), 1.94-1.86 (m, 1H), 1.80-1.73 (m, 1H), 1.58-1.46 (m, 2H), 1.42-1.29 (m, 4H), 1.29-1.19 (m, 2H), 1.01 (d, J = 18.1 Hz, 9H).

[0468] <Preparation Example 14> Preparation of Intermediate 2b (Int. 2.b) and Intermediate 2c (Int. 2.c)

[0469] Intermediate 2b of the present invention was prepared using the following reaction formulas 15 and 16, and intermediate 2c was prepared using the following reaction formulas 17 and 18.

[0470] [Reaction Equation 15]

[0471]

[0472] [Reaction Equation 16]

[0473]

[0474] [Reaction Equation 17]

[0475]

[0476] [Reaction Equation 18]

[0477]

[0478] 14-1. Preparation of Compound b.vi (tert-butyl ((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl) methyl)carbamate)

[0479] Compound b (1 eq), tert-butyl(piperidin-4-yl)methyl carbamate (1 eq), and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Subsequently, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex=1:1 solvent to obtain compound b.vi in ​​a yield of 58%.

[0480] 1 H NMR (600 MHz, DMSO) δ 11.06 (s, 1H), 7.63 (d, J = 8.5 Hz, 1H), 7.29 (d, J = 2.4 Hz, 1H), 7.23-7.19 (m, 1H), 6.88 (t, J = 6.0 Hz, 1H), 5.06-5.02 (m, 1H), 4.05-4.02 (m, 1H), 2.95-2.79 (m, 6H), 2.60-2.50 (m, 2H), 2.02-1.98 (m, 1H), 1.71-1.60 (m, 3H), 1.36 (s, 9H), 1.14-1.08 (d, J = 13.7 Hz, 2H).

[0481] 14-2. Preparation of Compound b.vii (tert-butyl (2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl) ethyl)carbamate)

[0482] Compound b (1 eq), tert-butyl 2-(piperidin-4-yl)ethyl carbamate (1 eq), and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Subsequently, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex=1:1 solvent to obtain compound b.vii in a yield of 52%.

[0483] 1 H NMR (600 MHz, DMSO) δ 11.06 (s, 1H), 7.63 (d, J = 8.6 Hz, 1H), 7.28 (d, J = 2.4 Hz, 1H), 7.23-7.19 (m, 1H), 6.77 (t, J = 5.8 Hz, 1H), 5.06-5.01 (m, 1H), 4.05-4.02 (m, 1H), 2.99-2.82 (m, 6H), 2.59-2.51 (m, 2H), 2.02-1.98 (m, 1H), 1.75-1.70 (m, 2H), 1.58-1.50 (m, 1H), 1.36 (s, 9H), 1.33-1.27 (m, 2H), 1.15-1.08 (m, 2H).

[0484] 14-3. Preparation of compound b.viii (tert-butyl (3-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperazin-1-yl)propyl)carbamate)

[0485] Compound b (1 eq), tert-butyl 3-(piperidin-4-yl)propyl carbamate (1 eq), and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Subsequently, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex=1:1 solvent to obtain compound b.viii in a yield of 57%.

[0486] 1 H NMR (600 MHz, DMSO) δ 11.07 (s, 1H), 7.66 (d, J = 8.5 Hz, 1H), 7.33 (d, J = 2.4 Hz, 1H), 7.26-7.22 (m, 1H), 6.80 (t, J = 5.7 Hz, 1H), 5.08-5.03 (m, 1H), 3.42 (t, J = 5.0 Hz, 4H), 2.97-2.91 (m, 2H), 2.91-2.82 (m, 1H), 2.61-2.51 (m, 2H), 2.46 (t, J = 5.1 Hz, 4H), 2.30 (t, J = 7.2 Hz, 2H), 2.03-1.98 (m, 1H), 1.58-1.51 (m, 2H), 1.36 (s, 9H).

[0487] 14-4. Preparation of Compound b.ix (tert-butyl ((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl) methyl)carbamate)

[0488] Compound b (1 eq), tert-butyl(azetidine-3-yl)methyl carbamate (1 eq), and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Subsequently, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex=1:1 solvent to obtain compound b.ix in a yield of 51%.

[0489] 1 H NMR (600 MHz, DMSO) δ 11.07 (s, 1H), 7.64 (d, J = 8.3 Hz, 1H), 7.09 (t, J = 5.9 Hz, 1H), 6.75 (d, J = 2.1 Hz, 1H), 6.65-6.59 (m, 1H), 5.08-5.02 (m, 1H), 4.08-4.04 (m, 2H), 3.74-3.68 (m, 2H), 3.21 (t, J = 6.3 Hz, 2H), 2.92-2.82 (m, 2H), 2.62-2.52 (m, 2H), 2.04-1.99 (m, 1H), 1.38 (s, 9H).

[0490] 14-5. Preparation of Compound bx (tert-butyl (2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl) ethyl)carbamate)

[0491] Compound b (1 eq), tert-butyl 2-(azetidine-3-yl)ethyl carbamate (1 eq), and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Subsequently, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was purified by silica column chromatography using EA:Hex=1:1 solvent to obtain compound bx in a yield of 46%.

[0492] 1 H NMR (600 MHz, DMSO) δ 11.07 (s, 1H), 7.63 (d, J = 8.3 Hz, 1H), 6.86 (t, J = 5.8 Hz, 1H), 6.75 (d, J = 2.1 Hz, 1H), 6.64-6.59 (m, 1H), 5.07-5.02 (m, 1H), 4.12 (t, J = 8.1 Hz, 2H), 3.68-3.62 (m, 2H), 2.97-2.91 (m, 2H), 2.91-2.83 (m, 1H), 2.80-2.72 (m, 1H), 2.62-2.52 (m, 2H), 2.04-2.00 (m, 1H), 1.77-1.71 (m, 2H), 1.39 (s, 9H).

[0493] 14-6. Preparation of compound c.vi (tert-butyl ((1-(2-(2-methylene-6-oxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)methyl)carbamate)

[0494] Compound c (1 eq), tert-butyl(piperidin-4-yl)methyl carbamate (1 eq) and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Afterward, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was first purified by silica column chromatography using EA:Hex=1:1 solvent and then used immediately for the next reaction.

[0495] 14-7. Compound c.vii (tert-butyl (2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)ethyl)carbamate) preparation

[0496] Compound c (1 eq), tert-butyl 2-(piperidin-4-yl)ethyl carbamate (1 eq) and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Afterward, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was first purified by silica column chromatography using EA:Hex=1:1 solvent and then used immediately for the next reaction.

[0497] 14-8. Preparation of Compound c.viii (tert-butyl (3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)propyl)carbamate)

[0498] Compound c (1 eq), tert-butyl 3-(piperidin-4-yl)propyl carbamate (1 eq) and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Afterward, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was first purified by silica column chromatography using EA:Hex=1:1 solvent and used immediately for the next reaction.

[0499] 14-9. Preparation of Compound c.ix (tert-butyl ((1-(2-(2-methylene-6-oxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)azetidin-3-yl)methyl)carbamate)

[0500] Compound c (1 eq), tert-butyl(azetidine-3-yl)methyl carbamate (1 eq) and DIPEA (2 eq) were mixed in DMSO and microwaved at 130 °C for 1 hour. Afterward, ethyl acetate and water were added to the mixture, the organic layer was dried with MgSO4, filtered, and concentrated under vacuum. The residue was first purified by silica column chromatography using EA:Hex=1:1 solvent and used immediately for the next reaction.

[0501] 14-10. Preparation of Intermediate 2

[0502] Compounds b.vi, b.vii, b.viii, b.ix, bx, c.vi, c.vii, c.viii, or c.ix were each reacted in a 25% TFA / DCM solution at room temperature for 1 hour. Afterward, volatile components were removed under reduced pressure, and the obtained intermediates Int. 2.b.vi, Int. 2.b.vii, Int. 2.b.viii, Int. 2.b.ix, Int. 2.bx, Int. 2.c.vi, Int. 2.c.vii, Int. 2.c.viii, and Int. 2.c.ix were used directly in the next reaction step as crude materials.

[0503] <Preparation Example 15> Preparation of PROTAC-based compound (6)

[0504] The PROTAC-based compounds SD-2920 to 2929 of the present invention were prepared using the following reaction schemes 19 and 20.

[0505] [Reaction Equation 19]

[0506]

[0507] [Reaction Equation 20]

[0508]

[0509] 15-1. Preparation of Compound 12

[0510] LiOH·H2O (2 eq) was added to a solution of compound 8 (1 eq) dissolved in a THF / H2O (1:1) mixed solvent, and the mixture was stirred at room temperature for 1-2 hours. The acidic crude residue obtained after concentrating the reaction mixture was used immediately in the next reaction step.

[0511] Amine crude compounds Int. 2.b.vi, Int. 2.b.vii, Int. 2.b.viii, Int. 2.b.ix, Int. 2.bx, Int. 2.c.vi, Int. 2.c.vii, Int. 2.c.viii, or Int. 2.c.ix (1 eq) were each mixed with the acidic crude compound of compound 8 (1 eq), HATU (1.5 eq), and DIPEA (3 eq), dissolved in DMF, and stirred overnight at room temperature. After the reaction was complete, the reaction mixture was concentrated, and the residue was mixed with ethyl acetate and water. The organic layer was extracted, dried with MgSO4, filtered, and concentrated under reduced pressure. The residue was purified twice using normal-phase silica column chromatography (solvent: MeOH / MC=1:15) and reverse-phase silica column chromatography (solvent: H2O / MeOH) to obtain compounds 12h, 12i, 12j, 12k, 12l, 12m, 12n, 12o, and 12p, which were each used immediately in the following reactions.

[0512] 15-2. Compound SD-2920 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)piperidin-4-yl) methyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) manufacturing

[0513] A DCM solution containing compound 12h (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2920 in a yield of about 30%.

[0514] 1H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.27-8.20 (m, 1H), 8.04-7.95 (m, 1H), 7.68-7.61 (m, 1H), 7.59-7.51 (m, 3H), 7.46-7.38 (m, 2H), 7.36 (t, J = 7.7 Hz, 1H), 7.33-7.19 (m, 5H), 7.06-6.99 (m, 1H), 5.09-5.04 (m, 1H), 4.84-4.75 (m, 1H), 4.70 (t, J = 9.2 Hz, 1H), 4.12-3.98 (m, 3H), 3.97-3.92 (m, 1H), 3.85-3.78 (m, 1H), 3.70-3.64 (m, 1H), 3.60-3.53 (m, 1H), 3.42-3.35 (m, 1H), 3.04-2.93 (m, 3H), 2.90-2.80 (m, 4H), 2.62-2.55 (m, 1H), 2.49-2.42 (m, 1H), 2.30-2.21 (m, 1H), 2.14-2.05 (m, 1H), 2.04-1.98 (m, 1H), 1.97-1.77 (m, 2H), 1.76-1.63 (m, 3H), 1.24-1.09 (m, 2H), 1.04 (d, J = 18.1 Hz, 9H).

[0515] 15-3. 화합물 SD-2921 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-5-yl)piperidin-4-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0516] A DCM solution containing compound 12i (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2921 in a yield of about 21%.

[0517] 1H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.28-8.21 (m, 1H), 8.00-7.88 (m, 1H), 7.68-7.61 (m, 1H), 7.59-7.51 (m, 3H), 7.46-7.38 (m, 2H), 7.35 (t, J = 7.6 Hz, 1H), 7.33-7.17 (m, 5H), 7.06-7.00 (m, 1H), 5.10-5.02 (m, 1H), 4.84-4.74 (m, 1H), 4.69 (t, J = 9.7 Hz, 1H), 4.08-3.98 (m, 2H), 3.96-3.90 (m, 1H), 3.85-3.77 (m, 1H), 3.70-3.62 (m, 1H), 3.59-3.53 (m, 1H), 3.42-3.35 (m, 1H), 3.35-3.28 (m, 1H), 3.17-3.02 (m, 3H), 2.98-2.83 (m, 4H), 2.63-2.55 (m, 1H), 2.48-2.40 (m, 1H), 2.28-2.20 (m, 1H), 2.14-2.05 (m, 1H), 2.05-1.98 (m, 1H), 1.97-1.78 (m, 2H), 1.77-1.66 (m, 3H), 1.53-1.45 (m, 1H), 1.37-1.29 (m, 1H), 1.26-1.09 (m, 2H), 1.04 (d, J = 17.6 Hz, 9H).

[0518] 15-4. 화합물 SD-2922 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-5-yl)piperazin-1-yl)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan -2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0519] A DCM solution containing compound 12j (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2922 in a yield of about 24%.

[0520] 1H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.27-8.18 (m, 1H), 8.17-7.98 (m, 1H), 7.70-7.65 (m, 1H), 7.57-7.47 (m, 3H), 7.42-7.35 (m, 2H), 7.32 (t, J = 7.6 Hz, 1H), 7.28-7.17 (m, 5H), 7.03-6.94 (m, 1H), 5.09-5.02 (m, 1H), 4.79-4.71 (m, 2H), 4.66 (t, J = 8.6 Hz, 1H), 3.95-3.89 (m, 1H), 3.88-3.83 (m, 1H), 3.82-3.74 (m, 1H), 3.68-3.60 (m, 2H), 3.56-3.49 (m, 1H), 3.42-3.36 (m, 1H), 3.08-2.96 (m, 3H), 2.95-2.77 (m, 5H), 2.73-2.62 (m, 1H), 2.61-2.54 (m, 1H), 2.45-2.38 (m, 1H), 2.36-2.29 (m, 1H), 2.29-2.18 (m, 2H), 2.10-1.98 (m, 2H), 1.95-1.83 (m, 2H), 1.82-1.72 (m, 1H), 1.72-1.58 (m, 3H), 1.02 (d, J = 12.4 Hz, 9H).

[0521] 15-5. 화합물 SD-2923 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-5-yl)azetidin-3-yl)methyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0522] A DCM solution containing compound 12n (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)methyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2923 in a yield of about 34%.

[0523] 1H NMR (600 MHz, DMSO) δ 11.06 (s, 1H), 8.20 (d, J = 9.0 Hz, 1H), 8.16-8.05 (m, 1H), 7.60-7.47 (m, 5H), 7.43-7.35 (m, 1H), 7.34-7.30 (m, 1H), 7.28-7.18 (m, 3H), 7.03-6.94 (m, 2H), 6.93-6.89 (m, 1H), 6.89-6.82 (m, 1H), 5.05-4.99 (m, 1H), 4.81-4.74 (m, 2H), 4.66 (d, J = 8.9 Hz, 1H), 3.94-3.88 (m, 1H), 3.87-3.78 (m, 1H), 3.78-3.63 (m, 4H), 3.27-3.16 (m, 4H), 2.95-2.78 (m, 1H), 2.60-2.54 (m, 1H), 2.48-2.43 (m, 1H), 2.42-2.37 (m, 1H), 2.35-2.28 (m, 1H), 2.26-2.19 (m, 1H), 2.19-2.11 (m, 1H), 2.10-2.02 (m, 1H), 2.02-1.94 (m, 1H), 1.93-1.85 (m, 1H), 1.83-1.72 (m, 1H), 1.70-1.62 (m, 1H), 1.01 (d, J = 16.0 Hz, 9H).

[0524] 15-6. 화합물 SD-2924 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-5-yl)azetidin-3-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0525] A DCM solution containing compound 12o (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2924 in a yield of about 31%.

[0526] 1H NMR (600 MHz, DMSO) δ 11.06 (s, 1H), 8.21 (d, J = 8.4 Hz, 1H), 8.06-7.94 (m, 1H), 7.58-7.45 (m, 5H), 7.44-7.35 (m, 1H), 7.32 (t, J = 7.5 Hz, 1H), 7.28-7.17 (m, 3H), 7.02-6.94 (m, 2H), 6.93-6.90 (m, 1H), 6.89-6.83 (m, 1H), 5.05-4.99 (m, 1H), 4.82-4.72 (m, 2H), 4.69-4.63 (m, 1H), 3.95-3.88 (m, 1H), 3.88-3.83 (m, 1H), 3.82-3.59 (m, 4H), 3.22-3.08 (m, 4H), 2.91-2.80 (m, 1H), 2.59-2.54 (m, 1H), 2.47-2.42 (m, 1H), 2.42-2.28 (m, 1H), 2.27-2.15 (m, 2H), 2.13-2.01 (m, 2H), 2.01-1.94 (m, 1H), 1.93-1.83 (m, 1H), 1.79-1.69 (m, 1H), 1.69-1.60 (m, 1H), 1.57-1.46 (m, 2H), 1.01 (d, J = 15.4 Hz, 9H).

[0527] 15-7. 화합물 SD-2925 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoiso indolin-4-yl)piperidin-4-yl)methyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0528] A DCM solution containing compound 12k (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)methyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2925 in a yield of about 22%.

[0529] 1H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.25-8.17 (m, 1H), 8.04-7.94 (m, 1H), 7.70-7.62 (m, 2H), 7.56-7.47 (m, 3H), 7.44-7.36 (m, 2H), 7.36-7.28 (m, 3H), 7.27-7.17 (m, 3H), 7.03-6.96 (m, 1H), 5.09-5.03 (m, 1H), 4.82-4.73 (m, 2H), 4.70-4.63 (m, 1H), 3.96-3.89 (m, 1H), 3.84-3.77 (m, 1H), 3.72-3.60 (m, 3H), 3.52-3.59 (m, 1H) 3.41-3.36 (m, 1H), 3.09-3.02 (m, 2H), 3.00-2.94 (m, 1H), 2.93-2.75 (m, 4H), 2.61-2.54 (m, 1H), 2.47-2.43 (m, 1H), 2.29-2.19 (m, 1H), 2.12-2.05 (m, 1H), 2.04-1.96 (m, 1H), 1.95-1.85 (m, 1H), 1.81-1.65 (m, 3H), 1.49-1.60 (m, 1H), 1.40-1.25 (m, 2H), 1.02 (d, J = 18.6 Hz, 9H).

[0530] 15-8. 화합물 SD-2926 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-4-yl)piperidin-4-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2 -yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0531] A DCM solution containing compound 12l (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0°C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0°C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2926 in a yield of about 23%.

[0532] 1H NMR (600 MHz, DMSO) δ 11.08 (s, 1H), 8.23-8.16 (m, 1H), 7.97-7.86 (m, 1H), 7.70-7.62 (m, 2H), 7.56-7.47 (m, 3H), 7.44-7.36 (m, 2H), 7.36-7.28 (m, 3H), 7.28-7.18 (m, 3H), 7.03-6.96 (m, 1H), 5.10-5.04 (m, 1H), 4.82-4.73 (m, 2H), 4.70-4.63 (m, 1H), 3.96-3.89 (m, 1H), 3.84-3.77 (m, 1H), 3.70-3.61 (m, 3H), 3.52-3.59 (m, 1H) 3.41-3.36 (m, 1H), 3.19-3.13 (m, 2H), 3.12-3.02 (m, 1H), 2.93-2.75 (m, 4H), 2.60-2.55 (m, 1H), 2.47-2.40 (m, 1H), 2.27-2.18 (m, 1H), 2.12-2.05 (m, 1H), 2.04-1.96 (m, 1H), 1.93-1.85 (m, 1H), 1.81-1.70 (m, 3H), 1.58-1.50 (m, 1H), 1.50-1.43 (m, 1H), 1.40-1.34 (m, 2H), 1.33-1.25 (m, 1H), 1.02 (d, J = 17.7 Hz, 9H).

[0533] 15-9. 화합물 SD-2927 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-4-yl)piperazin-1-yl)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan -2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0534] A DCM solution containing compound 12m (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((3-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)propyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0°C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0°C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2927 in a yield of about 33%.

[0535] 1H NMR (600 MHz, DMSO) δ 11.09 (s, 1H), 8.32-8.15 (m, 2H), 7.74-7.67 (m, 1H), 7.57-7.48 (m, 4H), 7.44-7.34 (m, 3H), 7.34-7.30 (m, 1H), 7.30-7.18 (m, 4H), 7.03-6.94 (m, 1H), 5.11-5.04 (m, 1H), 4.81-4.70 (m, 2H), 4.70-4.62 (m, 1H), 3.96-3.89 (m, 1H), 3.89-3.83 (m, 1H), 3.81-3.74 (m, 1H), 3.68-3.60 (m, 2H), 3.55-3.52 (m, 1H), 3.44-3.39 (m, 1H), 3.12-2.97 (m, 4H), 2.94-2.81 (m, 4H), 2.80-2.74 (m, 1H), 2.61-2.55 (m, 1H), 2.47-2.39 (m, 1H), 2.38-2.28 (m, 1H), 2.27-2.18 (m, 1H), 2.12-1.97 (m, 3H), 1.96-1.84 (m, 2H), 1.83-1.63 (m, 4H), 1.02 (d, J = 10.1 Hz, 9H).

[0536] 15-10. 화합물 SD-2929 (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxo isoindolin-4-yl)azetidin-3-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) 제조

[0537] A DCM solution containing compound 12p (((4-((E)-3-(((2S)-1-((2S)-2-(benzyl(3-((2-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)azetidin-3-yl)ethyl)amino)-3-oxopropyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)amino)-3-oxoprop-1-en-1-yl)phenyl)difluoromethyl)phosphonic acid) (1 eq) was cooled to 0 °C in an ice bath, and then CF3CON(TMS)2 (6 eq) and TMS-I (5 eq) were added to the reaction solution. The reaction mixture was stirred at 0 °C for 15-30 minutes, and then the solvent was removed under vacuum. The residue was purified with H2O / MeOH solvent using reverse-phase silica column chromatography to obtain compound SD-2929 in a yield of about 30%.

[0538] 1H NMR (600 MHz, DMSO) δ 11.10 (s, 1H), 8.24-8.15 (m, 1H), 8.04-7.91 (m, 1H), 7.58-7.45 (m, 4H), 7.44-7.35 (m, 2H), 7.34-7.28 (m, 1H), 7.28-7.17 (m, 3H), 7.16-7.09 (m, 1H), 7.04-6.96 (m, 2H), 6.64-6.56 (m, 1H), 5.08-4.99 (m, 1H), 4.81-4.72 (m, 2H), 4.70-4.62 (m, 1H), 3.95-3.89 (m, 1H), 3.89-3.82 (m, 1H), 3.81-3.74 (m, 1H), 3.69-3.59 (m, 2H), 3.22-3.16 (m, 1H), 3.13-3.02 (m, 2H), 2.97-2.91 (m, 1H), 2.90-2.80 (m, 2H), 2.61-2.53 (m, 1H), 2.47-2.41 (m, 1H), 2.41-2.34 (m, 1H), 2.26-2.17 (m, 1H), 2.12-1.95 (m, 2H), 1.95-1.79 (m, 2H), 1.78-1.69 (m, 1H), 1.69-1.62 (m, 1H), 1.61-1.44 (m, 2H), 1.42-1.32 (m, 1H), 1.02 (d, J = 16.7 Hz, 9H).

[0539] <Example 1> Confirmation of STAT6 degradation effect of STAT6-PROTAC compound (in vitro)

[0540] <1-1> Verification of STAT6 Decomposition Effect

[0541] We confirmed whether the STAT6-PROTAC compound of the present invention reduces the protein amount by degrading STAT6 (Signal transducer and activator of transcription 6) in human dermal fibroblasts (HDF). Specifically, 1 × 10⁶ HDF cells were placed in a 6-well plate. 5Cells were inoculated at a cell concentration, and 26 of the novel STAT6-PROTAC compounds synthesized in Preparation Examples 1 to 35 were selected and each treated at a concentration of 10 μM, and cultured for 18 hours. After the end of culture, cells were obtained, and proteins were extracted by treating with 40 μl of RIPA buffer. The amount of STAT6 protein was confirmed by Western blot and quantified by GAPDH. The results are shown in Table 2 and Figure 1 below.

[0542] STAT6-PROTAC drug PROTAC structure STAT6 Inhibition rate (%)SD-2744B-1-C50.5432SD-2745B-1-C11.4513SD-2746B-1-C18.4514SD-2780B-1-C-5.0395SD-2781B-1-C6.5594SD-2782B-1-C58.2717SD-27 83B-1-C11.0121SD-2875B-1-C31.5208SD-2876B-1-C46.2147SD-2877B-1 -C80.2024SD-2902B-1-C75.1144SD-2909B-1-C70.9277SD-2920B-1-C85.4 115SD-2921B-1-C85.6802SD-2922B-1-C94.7619SD-2923B-1-C85.1508SD -2924B-1-C80.2271SD-2925B-1-C86.1836SD-2926B-1-C88.5947SD-2927 B-1-C86.2209SD-2929B-1-C81.7927SD-2834B-2-C25.6867SD-2835B-2-C 5.3698SD-2836B-2-C11.6255SD-2837B-2-C18.0274SD-2842B-2-C64.9754

[0543] As a result, as shown in Table 1 and Figure 1 above, it was confirmed that some of the STAT6-PROTAC compounds of the present invention have excellent STAT6 degradation rates. In particular, it was confirmed that the compounds SD-2744, SD-2782, SD-2877, SD-2902, SD-2909, SD-2920, SD-2921, SD-2922, SD-2923, SD-2924, SD-2925, SD-2926, SD-2927, SD2929, and SD-2842 have STAT6 degradation rates of 50% or more.

[0544] <1-2> Confirmation of STAT6-specific degradation effect

[0545] It was confirmed whether compounds SD-2744, SD-2782, and SD-2877 among the STAT6-PROTAC compounds of the present invention specifically degrade only STAT6 among STATs. Specifically, 1 × 10⁶ HDF cells were placed in a 6-well plate. 5 Cells were inoculated at a certain concentration, treated with compounds SD-2744, SD-2782, and SD-2877 at a concentration of 10 μM each, and cultured for 18 hours. After the end of culture, cells were harvested, proteins were extracted by treating with 40 μl of RIPA buffer, and the amounts of STAT6, STAT3, and STAT5 proteins were confirmed by Western blotting and quantified using GAPDH protein. In addition, splenocytes were cultured in the same manner as above, and the amount of STAT6 protein was quantified after 18 hours.

[0546] As a result, as shown in FIGS. 2 to 4, it was confirmed that the compounds SD-2744, SD-2782, and SD-2877 of the present invention specifically degrade only STAT6 among STATs, thereby reducing the amount of STAT6 protein. In addition, it was confirmed that the compounds SD-2744, SD-2782, and SD-2877 degrade STAT6 in spleen cells as well (Figs. 5 to 7).

[0547] <1-3> Confirmation of half-maximum inhibitory concentration

[0548] The half maximal inhibitory concentration (IC 50) of the STAT6-PROTAC compounds of the present invention against STAT6 was determined. Specifically, under the same conditions as in Example 1-1 above, compounds SD-2744, SD-2782, and SD-2877 were diluted tenfold from a concentration of 10 μM to a concentration of 1 nM and treated to HDF cells, and the amount of STAT6 protein was analyzed by Western blotting after 18 hours.

[0549] As a result, as shown in Figure 8, it was confirmed that the IC 50 value of SD-2744 was 100 nM or less, the IC 50 value of SD-2782 was 0.1 nM, and the IC 50 value of SD-2877 was 0.5~0.6 nM.

[0550] <Example 2> Confirmation of improvement of systemic sclerosis by STAT6-PROTAC compound (in vivo)

[0551] <2-1> Confirmation of Inhibition of Tissue Fibrosis in Systemic Sclerosis

[0552] We confirmed whether the STAT6-PROTAC compound of the present invention inhibits tissue fibrosis in an animal model of systemic sclerosis. Specifically, an animal model in which systemic sclerosis was hyperactivated by immunosensitization with vimentin protein was constructed. At the start of the experiment, SKG mice were immunized by administering vimentin protein as an antigen along with Freund's complete adjuvant, followed by daily subcutaneous injection of 100 μl of bleomycin (BLM) for 3 weeks to induce systemic sclerosis. Additionally, at the second week of the experiment, the mice were further immunized using vimentin protein, and SD-2744, SD-2782, or SD-2877 were intravenously injected three times a week at a concentration of 00 M / kg for 3 weeks. The specific experimental procedure and drug administration schedule are shown in Figure 9. At the end of the experiment, the mice were humanely sacrificed, and skin and lung tissues were obtained and sectioned. Dermal thickness and lung fibrosis score were quantified using hematoxylin-eosin (H&E) staining of the sectioned tissues, and collagen within the tissue was quantified using Masson's Trichrome (MT) staining to confirm whether tissue fibrosis was inhibited. As a control, a vehicle group treated with an equal amount of solvent was used in a vimentin-sensitized systemic sclerosis (BLM+Vim) animal model.

[0553] As a result, as shown in FIGS. 10 to 12, compared to the control group, the dermal thickness and lung fibrosis score were significantly reduced in each group of mice treated with SD-2744, SD-2782, or SD-2877 of the present invention, and it was confirmed that collagen in the skin and lung tissues was significantly reduced, thereby confirming that the novel STAT6-PROTAC compound of the present invention inhibits tissue fibrosis in systemic sclerosis.

[0554] <2-2> Confirmation of Cytokine Inhibition and STAT6 Degradation in Systemic Sclerosis Fibrosis

[0555] It was confirmed whether the STAT6-PROTAC compound of the present invention degrades IL-4, a cytokine that specifically induces skin fibrosis in systemic sclerosis, and STAT6 within the tissue. Specifically, the amounts of IL-4 and STAT6 within the tissue were quantified using immunohistochemical staining on the skin tissue sectioned in Example 2-1. In addition, the amount of STAT6 within the tissue was confirmed using immunohistochemical staining on the lung tissue sectioned.

[0556] As a result, as shown in Figures 13 and 14, it was confirmed that the amount of IL-4, a systemic sclerosis fibrosis-associated cytokine, was significantly reduced in the skin tissue of the group administered the SD-2744 or SD-2877 compounds of the present invention compared to the control group. In addition, it was confirmed that the amount of STAT6 in the skin tissue was also significantly reduced by the administration of the SD-2744 or SD-2877 compounds.

[0557] In addition, as shown in FIGS. 15 to 17, the amount of STAT6 in lung tissue was checked, and compared to the control group, it was confirmed that the amount of STAT6 in lung tissue was significantly reduced in the group administered the compounds SD-2744, SD2782, or SD-2877 of the present invention, confirming that the STAT6-PROTAC compounds of the present invention inhibit cytokines that induce tissue fibrosis and degrade STAT6 in tissue.

[0558] <2-3> Confirmation of Fibrosis Factor Inhibition and STAT6 Degradation in Lung Tissue

[0559] It was confirmed whether the STAT6-PROTAC compound of the present invention reduces fibrosis factors in lung tissue and actually degrades STAT6 protein. Specifically, the lung tissues of each group obtained in Example 2-1 were crushed, and proteins were extracted using RIPA buffer. Subsequently, the protein amounts of the fibrosis factor α-smooth muscle actin (α-SMA) and STAT6 were confirmed by Western blotting, and quantified using the amount of GAPDH protein.

[0560] As a result, as shown in Figures 18 and 19, it was confirmed that the protein expression of α-SMA, a fibrosis factor, was significantly suppressed in the lung tissue of the group administered SD-2744 of the present invention compared to the control group. In addition, it was confirmed that the amount of STAT6 protein in the lung tissue was also significantly reduced by the administration of SD-2744 or SD-2877.

[0561] Accordingly, the present invention synthesized novel STAT6-PROTAC compounds capable of specifically degrading STAT6, and confirmed that the synthesized group of STAT6-PROTAC compounds specifically inhibits STAT6 in skin fibroblasts. Furthermore, it was confirmed that the STAT6-PROTAC compounds of the present invention inhibit fibrosis in skin and lung tissues in an animal model of systemic sclerosis sensitized with vimentin to increase pathological activity. Additionally, it was confirmed that they improve systemic sclerosis by inhibiting the expression of skin fibrosis cytokines and fibrosis factors.

Claims

A chimeric compound represented by the following chemical formula 1, its isomer, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, X is a protein targeting moiety, and L is the linker, and Y is a ubiquitin ligase binding moiety, and The above protein targeting moiety is a compound represented by the following chemical formula A; [Chemical Formula A] In the above chemical formula A, R1 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a carboxyl group, or an ester group, and R2 is either selected from hydrogen or halogen, and R3 is any one selected from hydrogen, straight-chain or branched-chain C1-10 alkyl groups. In paragraph 1, The above protein targeting moiety is one that binds to the STAT6 (Signal Transducer And Activator Of Transcription 6) protein, Chimeric compounds, isomers thereof, or pharmaceutically acceptable salts thereof. In paragraph 1, The compound represented by the above chemical formula A is one selected from the group of compounds below, Chimeric compounds, isomers thereof, or pharmaceutically acceptable salts thereof: In paragraph 1, The above linker has the structure of the following chemical formula B, Chimeric compounds, isomers thereof, or pharmaceutically acceptable salts thereof: [Chemical Formula B] In the above chemical formula B, L1 is R4, or does not exist, The above R4 is a straight-chain or branched-chain C1-10 alkyl group, and L2 is any one selected from a substituted or unsubstituted 3-10 atom heterocyclic group comprising a straight-chain or branched C1-10 alkoxy group, a straight-chain or branched C1-10 alkylamine group, a straight-chain or branched C1-10 alkylamide group, a straight-chain or branched C1-10 alkenyl group, a C3-10 cycloalkyl group, a C3-10 aryloxy group, a C3-10 arylamine group, and one or more heteroatoms selected from N, O, S, Se, and Te. L1 is bonded to the compound of chemical formula A. In paragraph 1, The above linker is one selected from compounds represented by the following chemical formulas C to G, Chimeric compounds, isomers thereof, or pharmaceutically acceptable salts thereof: [Chemical Formula C] In the above chemical formula C, n is an integer from 0 to 10; [Chemical Formula D] In the above chemical formula D, n is an integer from 0 to 10; [Chemical Formula E] In the above chemical formula E, n is an integer from 0 to 10; [Chemical Formula F] In the above chemical formula F, n is an integer from 0 to 10; [Chemical Formula G] In the above chemical formula G, n is an integer from 0 to 10. In paragraph 1, The above ubiquitin ligase binding moiety is one that binds to E3 ubiquitin ligase, Chimeric compounds, isomers thereof, or pharmaceutically acceptable salts thereof. In paragraph 1, The above ubiquitin ligase binding moiety is optionally substituted thalidomide, pomalidomide, lenalidomide, prolinamide, a stereoisomer thereof, or a derivative thereof, Chimeric compounds, isomers thereof, or pharmaceutically acceptable salts thereof. In paragraph 1, The above ubiquitin ligase binding moiety is one selected from compounds represented by the following chemical formulas H to L, Chimeric compounds, isomers thereof, or pharmaceutically acceptable salts thereof: [Chemical Formula H] In the above chemical formula H, Z is H2 or O, and The above compound is bonded to L2 of chemical formula B. [Chemical Formula I] In the above chemical formula I, Z is H2 or O, and The above compound is bonded to L2 of chemical formula B. [Chemical Formula J] In the above chemical formula J, Z is H2 or O, and The above compound is bonded to L2 of chemical formula B. [Chemical Formula K] In the above chemical formula K, Z is H2 or O, and The above compound is bonded to L2 of chemical formula B. [Chemical Formula L] In the above chemical formula L, Z is H2 or O, and The above compound is bonded to L2 of chemical formula B. In paragraph 1, The above chimeric compound is a compound, an isomer thereof, or a pharmaceutically acceptable salt thereof, wherein the chimeric compound is any one selected from the compounds represented by the following chemical formulas 3 to 37. [Chemical Formula 3] [Chemical Formula 4] [Chemical Formula 5] [Chemical Formula 6] [Chemical Formula 7] [Chemical Formula 8] [Chemical Formula 9] [Chemical Formula 10] [Chemical Formula 11] [Chemical Formula 12] [Chemical Formula 13] [Chemical Formula 14] [Chemical Formula 15] [Chemical Formula 16] [Chemical Formula 17] [Chemical Formula 18] [Chemical Formula 19] [Chemical Formula 20] [Chemical Formula 21] [Chemical Formula 22] [Chemical Formula 23] [Chemical Formula 24] [Chemical Formula 25] [Chemical Formula 26] [Chemical Formula 27] [Chemical Formula 28] [Chemical Formula 29] [Chemical Formula 30] [Chemical Formula 31] [Chemical Formula 32] [Chemical Formula 33] [Chemical Formula 34] [Chemical Formula 35] [Chemical Formula 36] [Chemical Formula 37] A compound represented by the following chemical formula 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: [Chemical Formula 2] In the above chemical formula 2, R1 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a carboxyl group, or an ester group, and R2 is either selected from hydrogen or halogen, and R3 is any one selected from hydrogen, a straight-chain or branched-chain C1-10 alkyl group, and R5 is any one selected from hydrogen, halogen, a straight-chain or branched-chain C1-10 alkyl group, a straight-chain or branched-chain C1-10 alkoxy group, a straight-chain or branched-chain C1-10 alkylamine group, a carboxyl group, or an ester group. In Paragraph 10, The above compound is one selected from the group of compounds below, Compounds, isomers thereof, or pharmaceutically acceptable salts thereof: In Paragraph 11, The above compound targets STAT6, A compound, its isomer, or its pharmaceutically acceptable salt. A pharmaceutical composition for the prevention or treatment of systemic sclerosis (SSc) comprising a compound of either claim 1 or claim 10 as an active ingredient. In Paragraph 13, A composition in which the above compound targets STAT6 (Signal transducer and activator of transcription 6). In Paragraph 14, A composition that targets the above STAT6 and decomposes STAT6. In Paragraph 15, A composition in which the above compound inhibits tissue fibrosis. In Paragraph 16, A composition that inhibits the above tissue fibrosis, which reduces skin thickness. In Paragraph 16, A composition that inhibits the above tissue fibrosis, which reduces the pulmonary fibrosis index. In Paragraph 16, A composition that inhibits the above tissue fibrosis by reducing collagen in the tissue. In Paragraph 16, A composition that inhibits the above tissue fibrosis by inhibiting the expression of fibrosis cytokines or fibrosis factors. In Article 20, A composition in which the above-mentioned fibrotic cytokine is IL-4. In Article 20, A composition in which the above fibrotic factor is α-smooth muscle actin (α-SMA). In Paragraph 13, A composition in which the above compound reduces the amount of STAT6 protein in tissues. A method for treating systemic sclerosis (SSc), comprising the step of administering a pharmaceutically effective amount of the compound of either claim 1 or claim 10 to an individual.

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