1,3-isoindolindione derivatives and heteroaromatic analogs for stimulating endothelial cell-pericyte interactions

Abstract

The present invention relates to a pharmaceutical compound and a formulation comprising the compound, wherein the compound is of formula I or II: The present invention relates to a pharmaceutical compound and a formulation having the structure shown in JPEG2026522984000107.jpg75161. The compound and the formulation are suitable for pharmaceutical use in the prevention or treatment of diseases or disorders characterized by pericyte dysfunction, detachment and / or loss; in therapeutic treatments for vascular stabilization; in treatments for promoting, restoring or maintaining pericyte adhesion to blood vessels; and / or in the treatment of vascular-related diseases in patients.

Description

[Technical Field]

[0001] Field of Invention This invention relates to the field of pharmaceuticals. More specifically, it relates to the use of 1,3-isoindolindione derivatives and their heteroaromatic analogs for stimulating endothelial cell-pericyte interactions and vascular pericyte coating. [Background technology]

[0002] Background of the Invention Blood vessels are a complex network of hollow tubes that continuously adapt structurally and functionally to ensure optimal delivery of nutrients and oxygen to all cells in the body. They consist of endothelial cells, which form the inner lining of the vessel wall, and mural cells called vascular smooth muscle cells (VSMCs) and pericytes (PCs). VSMCs surround arteries and veins, while PCs are located within the basement membrane of capillaries, venules, and terminal arterioles. PCs play a crucial role in angiogenesis and stabilization, regulate endothelial cell survival, and control the blood-blood barrier and blood flow to solutes and immune cells. Many serious diseases affecting millions of people worldwide are caused by or related to PC death or migration from blood vessels. These pathological responses can lead to chronic inflammation, excessive vascular proliferation, or / or vascular dilution, which can ultimately result in hypoperfusion and tissue damage.

[0003] PC dysfunction has been reported to cause or contribute to the development of a wide range of conditions, including diabetes mellitus (nephropathy and retinopathy), chronic kidney disease (CKD), cardiomyopathy, central nervous system (CNS) disorders such as stroke, epilepsy, spinal cord injury, dementia, particularly Alzheimer's disease, Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis, radiation necrosis, and hereditary small vessel diseases such as autosomal dominant cerebral arteriovenous disease (Cadasil) and autosomal recessive cerebral arteriovenous disease (Carasil) with subcortical infarction and leukoencephalopathy, cerebral amyloid angiopathy (CAA), retinal vascular disorders with cerebral leukoencephalopathy and systemic symptoms (RVCL-S), hereditary hemorrhagic telangiectasia (HHT), and cavernous hemangioma (CCM). The majority of these conditions still lack the availability of effective treatments suitable for chronic management. We and others have previously investigated how promoting the adhesion of PC to blood vessels can improve HHT 1 by, or by radiation therapy to the brain 2-3 This provides proof-of-concept data suggesting that it can prevent vascular fragility caused by [the condition]. Thalidomide is particularly effective for this in both mice and patients, suggesting that it may be useful in other diseases with similar underlying pathologies.

[0004] Thalidomide and its analogs, lenalidomide and pomalidomide, are immunomodulatory imide drugs (IMiDs) and are highly effective treatments for multiple myeloma and del(5q) myelodysplastic syndrome. This class of compounds exhibits multifaceted effects, including immunomodulatory, potent anti-inflammatory, anti-angiogenic, and direct anti-myeloma activity. Mechanistically, these drugs act as molecular crosslinks that link selected neosubstrates to the ubiquitin-proteasome system for degradation. All IMiDs are characterized by a glutarimide ring that binds to cereblon (CRBN), a substrate of the E3 ubiquitin ligase CRL4 complex, which leads to the degradation of target proteins. The glutarimide moiety is responsible for most of their pharmacological activity, but also for their harmful teratogenic and hematological effects. In particular, accumulated evidence indicates that the immunomodulatory, anti-angiogenic, and direct antiproliferative activity of all IMiDs depends solely on the glutarimide motif, and not on the phthalimide motif.

[0005] IMiDs cannot be used to treat chronic diseases. Patients treated with IMiDs may develop serious side effects, including the possibility of birth defects, fatigue, weakness, anemia, neurological disorders, neutropenia, thrombocytopenia, thrombosis leading to stroke or heart attack, increased risk of death in patients with chronic lymphocytic leukemia (CLL), risk of malignancy, severe liver damage, severe skin reactions, thyroid disorders, and increased risk of premature death in patients with mantle cell lymphoma (MCL).

[0006] Therefore, an object of the present invention is to provide compounds and / or treatments for promoting endothelial cell-pericyte interactions and vascular pericyte coating, which can be used in chronic treatments, unlike currently known IMiDs. [Overview of the Initiative]

[0007] Therefore, the present invention relates to formula I or II: [ka] 〔In the formula, A 1 、A 2 、A 3 and A 4 each independently represents CR 1 or N; A 5 and A 6 each independently represents S, CR 1 or N; R ... 1 represents hydrogen, halogen, OH, C1-C8 alkyl, NO2, or NR 2 R 3 ; R 2 and R 3 are each independently selected from H, C1-C8 alkyl, C1-C8-alkyl-carbonyl, and C1-C8-alkoxy-carbonyl; R x represents Y or CHR 4 R 5 ; Y represents a 4-membered, 5-membered or 6-membered heterocycle containing one, two or three heteroatoms, Y may optionally be substituted with one or more substituents selected from the group consisting of halide, saturated or unsaturated C1-C8 hydrocarbon, C1-C8-alkoxy-carbonyl, or C1-C8-alkyl-carbonyl, preferably acetyl or propionyl; R 4 represents H, C1-C8-alkyl, C1-C8-cycloalkyl, C1-C8-cycloalkylalkyl, heteroaryl, C1-C8-heteroaralkyl, C1-C8-heterocycloalkyl, or C1-C8-heterocycloalkylalkyl, R 5 represents H, OH, COOH, or C1-C8 alkyl, preferably COOH, i-propyl or t-butyl; provided that the case where both R 4 and R 5 are H is excluded〕 relates to a pharmaceutical compound having the structure shown in

[0008] References to the compounds of formulas (I) and (II) described herein, as well as their subreferences, also include pharmaceutically acceptable ionic forms, salts, solvates, isomers including geometric and stereoisomers, tautomers, N-oxides, esters, prodrugs, isotopes and their protected forms, preferably pharmaceutically acceptable salts or prodrugs.

[0009] In a second aspect, the present invention relates to a pharmaceutical use in therapeutic treatments for stimulating endothelial cell-pericyte interactions and promoting the vascular PC capsule, comprising formula I or II: [ka] [During the ceremony, A 1 , A 2 , A 3 and A 4 Each of them operates independently, CR 1 Or it represents N; A 5 and A 6 These are S and CR, which are independent of each other. 1 Or it represents N; R 1 This includes hydrogen, halogens, OH, C1-C8 alkyl, NO2, or NR 2 R 3 This represents, R 2 and R 3 Each of these is independently selected from H, C1-C8 alkyl, C1-C8 alkyl-carbonyl, and C1-C8 alkoxy-carbonyl; R x is H, Y, or CHR 4 R 5 This represents, Y represents a 4-membered, 5-membered, or 6-membered heteroring containing one, two, or three heteroatoms. Y may optionally be substituted with one or more substituents selected from the group consisting of halides, saturated or unsaturated C1-C8 hydrocarbons, C1-C8 alkoxycarbonyls, or C1-C8 alkylcarbonyls, preferably acetyl or propionyl; R 4 This represents H, C1~C8-alkyl, C1~C8-cycloalkyl, C1~C8-cycloalkylalkyl, heteroaryl, C1~C8-heteroaralkyl, C1~C8-heterocycloalkyl, or C1~C8-heterocycloalkylalkyl. R 5 represents H, OH, COOH, C1-C8 alkyl, preferably COOH, i-propyl or t-butyl; optionally, R 4 and R 5 [Except when both are H] This relates to a pharmaceutical compound having the structure shown.

[0010] In a third embodiment, the present invention relates to the use of a compound I or II for pharmaceutically acceptable treatment in the prophylactic or therapeutic treatment of diseases or disorders characterized by pericyte dysfunction, detachment, and / or loss: [ka] [During the ceremony, A 1 , A 2 , A 3 and A 4 Each of them operates independently, CR 1 Or it represents N; A 5 and A 6 These are S and CR, which are independent of each other. 1 Or it represents N; R 1 This includes hydrogen, halogens, OH, C1-C8 alkyl, NO2, or NR 2 R 3 This represents, R 2 and R 3Each of these is independently selected from H, C1-C8 alkyl, C1-C8 alkyl-carbonyl, and C1-C8 alkoxy-carbonyl; R x is H, Y, or CHR 4 R 5 This represents, Y represents a 4-membered, 5-membered, or 6-membered heteroring containing one, two, or three heteroatoms. Y may optionally be substituted with one or more substituents selected from the group consisting of halides, saturated or unsaturated C1-C8 hydrocarbons, C1-C8 alkoxycarbonyls, or C1-C8 alkylcarbonyls, preferably acetyl or propionyl; R 4 This represents H, C1~C8-alkyl, C1~C8-cycloalkyl, C1~C8-cycloalkylalkyl, heteroaryl, C1~C8-heteroaralkyl, C1~C8-heterocycloalkyl, or C1~C8-heterocycloalkylalkyl. R 5 represents H, OH, COOH, C1-C8 alkyl, preferably COOH, i-propyl or t-butyl; optionally, R 4 and R 5 [Except when both are H] This relates to a pharmaceutical compound having the structure shown. [Brief explanation of the drawing]

[0011] [Figure 1] Figure 1 shows the relative effectiveness of selected phthalimides in preventing radiotherapy-induced pericyte detachment and vascular leakage. [Figure 2] Figure 2 shows the relative effectiveness of selected phthalimides in preventing radiotherapy-induced delayed vascular dilution and neuronal loss in the somatosensory cortex (SS) and hippocampal region (Hp). [Figure 3] Figure 3 illustrates the vascular stabilization properties of selected phthalimides, as demonstrated using a mouse model of hereditary hemorrhagic telangiectasia (HHT) type 1. [Figure 4] Figure 4 shows selected analogues of phthalimide that restore endothelial cell-pericyte interactions and vascular stability according to one embodiment of the present invention. [Modes for carrying out the invention]

[0012] Detailed description of the invention The present invention provides novel 1,3-isoindolindione derivatives and their heteroaromatic analogs, as well as 1,3-isoindolindione derivatives and their heteroaromatic analogs for use in stimulating endothelial cell-PC interactions and promoting vascular PC capsule formation, and for use in the prevention or treatment of diseases or disorders characterized by pericyte dysfunction, pericyte detachment, and / or pericyte loss.

[0013] It was surprisingly discovered that these compounds can prevent pericyte (PC) detachment and promote endothelial cell-pericyte interactions and pericyte coverage of blood vessels. Therefore, the compounds of the present invention can be advantageously used in the prophylactic or therapeutic treatment of diseases or disorders characterized by pericyte dysfunction, detachment, and / or loss, including therapeutic treatments including promoting endothelial cell-PC interactions and vascular stabilization, as well as prophylactic and / or therapeutic treatments including promoting, restoring, or maintaining PC adhesion to blood vessels.

[0014] The inventors have found that thalidomide, unlike its analog lenalidomide, can prevent radiotherapy-induced pericyte (PC) detachment (Figure 1A-B) and blood-brain barrier (BBB) ​​disruption (Figure 1C) in mice, which serve as a model for early vascular injury associated with delayed neuronal loss and cognitive decline.

[0015] The inventors also found that prophylactic treatment with thalidomide, unlike treatment with its analogue lenalidomide (Figure 2A), can prevent delayed capillary dilution induced by radiotherapy in the cortical region (Figure 2B-F) or hippocampal region (Figure 2B, 2G-J) and prevent neuronal loss by preventing pericyte detachment and blood-brain barrier (BBB) ​​leakage. Furthermore, thalidomide and pomalidomide, in contrast to their analogue lenalidomide, were observed to stimulate endothelial cell-PC interactions and vascular coverage in a mouse model of HHT using ex vivo electrophysiological techniques to quantify endothelial cell-PC interactions (Figure 3). As a result, it was inferred that the carbonyl group (C=O) of the phthaloyl ring plays a crucial role in thalidomide's ability to promote pericyte recruitment to the endothelium. Furthermore, apremilast, an oral small molecule inhibitor of phosphodiesterase 4 (PDE4) that, like thalidomide, possesses a phthalimide ring but lacks a glutarimide ring and therefore cannot bind to cereblon, was found to be able to rescue endothelial cell-PC interactions in a mouse model of HHT at high concentrations. This confirmed that the glutarimide ring is not necessary to promote PC coating of the vascular system (Figure 3). Therefore, the isoindoline dione motif, found in some IMiDs such as thalidomide but not in lenalidomide, is considered to be the active fragment in compounds that promote PC adhesion to blood vessels. Moreover, it was found that the glutarimide motif, which is always present in IMiDs, is not essential for vascular stabilization, and that major side effects associated with known IMiDs can be favorably suppressed by removing or substituting this structural motif.

[0016] Furthermore, it was found that the claimed effect could be obtained not only with phthalimide derivatives containing a C6 aromatic moiety, but also with other aromatic moieties or heteroaromatic moieties.

[0017] Therefore, each A in the aromatic moiety of the 5-membered or 6-membered ring of the compound as defined herein 1 , A 2 , A 3and A 4 independently represents CR 1 or N, and A 5 and A 6 each independently represents S, CR 1 or N; R 1 is hydrogen, halogen, OH, C1-C8 alkyl, NO2, or NR 2 R 3 represents, and R 2 and R 3 are each independently selected from H, C1-C8 alkyl, C1-C8-alkyl-carbonyl, and C1-C8-alkoxy-carbonyl.

[0018] In certain embodiments, A 1 or A 2 represents C-NH2 or C-NHC(O)CH3; preferably, A 1 represents C-NH2, and A 2 , A 3 and A 4 each independently represents CH, C-CH3 or C-Cl, preferably CH. In certain embodiments, A 2 represents C-NH2, and A 1 , A 3 and A 4 each independently represents CH, C-CH3 or C-Cl, preferably CH.

[0019] In one embodiment, one of A 5 and A 6 is S, and the rest of A 4 , A 5 and A 6 are CR 1 or CH.

[0020] The compounds according to the present invention can be advantageously used in the treatment of diseases or disorders characterized by pericyte dysfunction, detachment, and / or loss, e.g., prophylactic and / or therapeutic treatments; these include therapeutic treatments that promote endothelial cell-PC interactions and vascular stabilization, e.g., therapeutic treatments, as well as prophylactic and / or therapeutic treatments that promote, restore, or maintain pericyte adhesion to blood vessels. In this specification, vascular stabilization means, among other things, protection of blood-BB integrity, control of blood flow, limitation of inflammation and hypoperfusion, and reduction of tissue damage, or a combination thereof.

[0021] While we do not wish to be bound by any particular theory, the major side effects of thalidomide and its analogs, lenalidomide and pomalidomide, are thought to be caused, at least in part, by their suitability and ability to bind to cereblon protein, which is the sole or primary target protein resulting in the degradation of multiple neosubstrates mediated by the CRL4CRBN ligase complex. Therefore, we believe that at least some of the major side effects of phthalimide derivatives are caused by the compound substantially preventing binding to cereblon protein. x We recognized that the toxicity could be suppressed by selecting the appropriate group. This finding enables rapid in vitro screening of the toxicity of compounds exhibiting the beneficial effects of the present invention.

[0022] The compounds defined herein are selected such that the compounds of the present invention do not bind to cereblon. x In the base, the maleimide moiety is N-substituted. This effectively excludes the glutarimide present in thalidomide and therefore thalidomide itself. In one embodiment, the lack of binding of the compound according to the present invention to cereblon (CRBN) compared to the binding of thalidomide to cereblon, as measured in a competitive binding assay, results in a maximum inhibitory concentration (IC) at half of the population. 50 This corresponds to a value higher than 100 μM, preferably higher than 200 μM, and more preferably higher than 500 μM or 1000 μM.

[0023] Thus, in certain embodiments, the compounds of Formula I do not include thalidomide, pomalidomide, lenalidomide or any other compounds containing a glutarimide moiety as present in these compounds.

[0024] R x is as defined herein and is further selected so as not to interfere with pharmaceutical activity.

[0025] In certain embodiments, R x is Y, where Y represents a 4-, 5- or 6-membered heterocycle containing one, two, or three heteroatoms. Preferably, the heteroatoms are independently selected from N, O, and S. Preferably, Y represents a 4-, 5-, or 6-membered heterocycle containing one heteroatom.

[0026] Particularly good results are obtained with compounds in which R x represents a substituted or unsubstituted 4-, 5- or 6-membered heterocycle such as a morpholine, azetidine, pyrrolidine or piperidine group.

[0027] Thus, in certain embodiments, R x represents a 4-, 5- or 6-membered heterocycle containing one, two, or three heteroatoms, preferably one heteroatom, preferably one nitrogen atom.

[0028] R x Azetidine as the group shows particularly good results. Thus, in a preferred embodiment of the invention, R x is

Chemical formula

[0029] Advantageously, these compounds have the corresponding amino acid H2N-C(R 8 It can be obtained relatively easily synthetically by the condensation reaction of 2-CO2H with the corresponding maleimide derivative.

[0030] In another embodiment, R x CHR 4 R 5 This represents, where R 4 This represents hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, tert-butyl, neopentyl, isopentyl, cyclopropyl-methyl, or thienyl-methyl.

[0031] One reason, R 5 This represents COOH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, tert-butyl, preferably COOH, methyl, i-propyl, or t-butyl.

[0032] In an exemplary embodiment, the compound is of formula (Ia) [ka] [In the formula, each Z is independently hydrogen, halogen, OH, C1-C8 alkyl, NO2, and NR] 2 R 3 Selected from, R 2 and R 3 Each of these is independently selected from H, C1-C8 alkyl, C1-C8-alkyl-carbonyl, and C1-C8-alkoxy-carbonyl, preferably each Z is independently selected from H, halogen, NH2, and NHC(O)CH3; m is 1, 2, 3, or 4, preferably 1 or 2; n is 1, 2, or 3, and R 6 [This represents saturated or unsaturated C1-C8 hydrocarbons, C1-C8 alkoxycarbonyls, or C1-C8 alkylcarbonyls, preferably acetyl or propionyl.] It has the structure shown.

[0033] In some exemplary embodiments, A 1 Or A 2 It is C-NH2, and R x is an N-substituted azetidine group. In preferred embodiments, the compound is of formula Ib or formula Ic: [ka] [In the formula, each Z is independently selected from hydrogen, halogen, OH, C1-C8 alkyl, and NO2, preferably each Z is independently selected from H, CH3, and Cl; m is 1, 2, or 3; and R 6 [This represents saturated or unsaturated C1-C8 hydrocarbons, C1-C8 alkoxycarbonyls, or C1-C8 alkylcarbonyls, preferably acetyl or propionyl.] It has the structure shown.

[0034] In another embodiment of the present invention, a formulation comprising a pharmaceutical compound as defined herein is provided, the formulation further comprising pharmaceutically acceptable additives, adjuvants, diluents and / or carriers.

[0035] Table 1 lists exemplary compounds according to the present invention. [Table 1-1] [Table 1-2] [Table 1-3]

[0036] Therefore, in one embodiment, compounds (1), (2), (3), (4), (5), (6), (7), (8), (9), (10), (11), (12), (13), (14), (15), (16), (17), (18), (19), (20), (21), (22), (23), (24), (25), (26), (27), (28), (29), (30), (31), (32), (33), (34), or (35) shown in Table 1 are provided. In another embodiment, a formulation, such as a pharmaceutical formulation, is provided, comprising compound (1), (2), (3), (4), (5), (6), (7), (8), (9), (10), (11), (12), (13), (14), (15), (16), (17), (18), (19), (20), (21), (22), (23), (24), (25), (26), (27), (28), (29), (30), (31), (32), (33), (34), or (35) shown in Table 1, along with pharmaceutically acceptable additives, adjuvants, diluents, and / or carriers. In another embodiment, compounds (1), (2), (3), (4), (5), (6), (7), (8), (9), (10), (11), (12), (13), (14), (15), (16), (17), (18), (19), (20), (21), (22), (23), (24), (25), (26), (27), (28), (29), (30), (31), (32), (33), (34), or (35) are provided for use as pharmaceuticals, in particular for preventive or therapeutic treatment of diseases or disorders characterized by pericyte dysfunction, detachment and / or loss, preferably for treatment to promote, restore or maintain pericyte adhesion to blood vessels. In one exemplary embodiment, compounds (1), (2), (5), (6), (7), (9), (10), (11), (15), (16), (17), (18), (19), (20), (21), (22), (23), or (24) shown in Table 1 are provided.In another embodiment, formulations comprising compound (1), (2), (5), (6), (7), (9), (10), (11), (15), (16), (17), (18), (19), (20), (21), (22), (23), or (24) with pharmaceutically acceptable additives, adjuvants, diluents and / or carriers, e.g., pharmaceutical formulations, are provided. In another exemplary embodiment, compounds (1), (2), (5), (6), (7), (9), (10), (11), (15), (16), (17), (18), (19), (20), (21), (22), (23), or (24) for pharmaceutically use, particularly for pharmaceutically use as defined herein.

[0037] Table 2 lists further exemplary compounds that conform to formulas Ia, Ib, and Ic. [Table 2-1] [Table 2-2] [Table 2-3]

[0038] Accordingly, in one embodiment, compounds 1.001 to 1.088 disclosed in Table 2 are provided. In another embodiment, formulations comprising one or more compounds 1.001 to 1.088 and pharmaceutically acceptable additives, adjuvants, diluents and / or carriers are provided, such as pharmaceutical formulations. In yet another embodiment, formulations comprising compounds 1.001 to 1.088, or one or more of these compounds and pharmaceutically acceptable additives, adjuvants, diluents and / or carriers, are provided for pharmaceutically appropriate use, particularly for the treatment of diseases or disorders characterized by pericyte dysfunction, detachment and / or loss, preferably for the treatment of promoting, restoring or maintaining pericyte adhesion to blood vessels.

[0039] Table 3 lists exemplary compounds that follow formula Ia. [Table 3-1] [Table 3-2] [Table 3-3]

[0040] Accordingly, in one embodiment, compounds 2.001 to 2.088 disclosed in Table 3 are provided. In another embodiment, a formulation is provided comprising one or more of compounds 2.001 to 2.088 disclosed in Table 3 and a pharmaceutically acceptable additive, adjuvant, diluent and / or carrier. In yet another embodiment, a formulation is provided comprising compounds 2.001 to 2.088, or one or more of these compounds and a pharmaceutically acceptable additive, adjuvant, diluent and / or carrier, for pharmaceutically appropriate use, particularly for the treatment of diseases or disorders characterized by pericyte dysfunction, detachment and / or loss, preferably for the treatment of promoting, restoring or maintaining pericyte adhesion to blood vessels.

[0041] Additives are natural or synthetic substances formulated together with the active ingredient (e.g., nucleic acid sequences, vectors, modified cells, or isolated peptides provided herein) for the purpose of increasing the volume of the formulation in the final dosage form or providing therapeutic enhancement to the active ingredient, such as by promoting drug absorption or solubility. Additives may also be useful in the manufacturing process to assist in the handling of the active ingredient by promoting powder flowability or non-stickiness, and may further be useful in assisting in vitro stability, such as preventing denaturation over the expected shelf life. Pharmaceutically acceptable additives are well known in the art. Therefore, suitable additives can be readily identified by those skilled in the art. Examples of suitable pharmaceutically acceptable additives include water, saline solution, aqueous dextrose, glycerol, and ethanol. Diluents are substances used for dilution. Pharmaceutically acceptable diluents are well known in the art. Therefore, suitable diluents can be readily identified by those skilled in the art. Carriers are nontoxic to the target at the dose and concentration used and are compatible with the other components of the formulation. The term "carrier" refers to an organic or inorganic component, natural or synthetic, that is combined with an active ingredient to facilitate application. Pharmacovigilantly acceptable carriers are well known in the art. Therefore, a suitable carrier can be readily identified by those skilled in the art. An adjuvant is a pharmacological and / or immunological agent that alters the effects of other substances in a formulation. Pharmacovigilantly acceptable adjuvants are well known in the art. Therefore, a suitable adjuvant can be readily identified by those skilled in the art.

[0042] The compounds of the present invention may be used in therapeutic treatments including prophylactic and therapeutic treatments for diseases or disorders characterized by pericyte dysfunction or pericyte loss, including therapeutic treatments including promoting endothelial cell-pericyte interactions and vascular stabilization, and prophylactic and / or therapeutic treatments including restoring, promoting or maintaining pericyte attachment to blood vessels.

[0043] In one embodiment, a disease or disorder characterized by pericyte dysfunction, detachment, and / or loss is selected from diabetic complications, chronic kidney disease (CKD), small vessel disease, and central nervous system (CNS) diseases.

[0044] In one embodiment, a disease or disorder characterized by pericyte dysfunction, pericyte detachment, and / or pericyte loss is selected from diabetic nephropathy (DN), diabetic retinopathy (DR), diabetic cardiomyopathy (DCM), and diabetic neuropathy.

[0045] In one embodiment, the disease or disorder characterized by pericyte dysfunction, detachment, and / or loss is selected from hypertensive nephropathy, IgA nephropathy, congenital nephrotic syndrome, lupus nephritis, polycystic kidney disease, and allograft nephropathy.

[0046] In one embodiment, a disease or disorder characterized by pericyte dysfunction, detachment, and / or loss is selected from autosomal dominant cerebral arteriopathies with subcortical infarction and leukoencephalopathy (Cadasil), autosomal recessive cerebral arteriopathies with subcortical infarction and leukoencephalopathy (Carasil), cerebral amyloid angiopathy (CAA), retinal vascular disorder with cerebral leukoencephalopathy and systemic symptoms (RVCL-S), hereditary hemorrhagic telangiectasia (HHT), and cavernous hemangioma (CCM).

[0047] In one embodiment, a disease or disorder characterized by pericyte dysfunction, detachment, and / or loss is selected from stroke, epilepsy, spinal cord injury, vascular dementia, Alzheimer's disease, Huntington's disease, Parkinson's disease, traumatic brain injury, multiple sclerosis, amyotrophic lateral sclerosis, and radiation necrosis.

[0048] In further embodiments, the present invention provides compounds or formulations as defined herein for use as pharmaceuticals to stimulate endothelial cell-pericyte interactions and promote vascular pericyte coating in patients, as well as methods for stimulating endothelial cell-PC interactions and promoting vascular PC coating in patients, comprising administering the compounds or formulations as defined herein.

[0049] In another embodiment, the present invention relates to compounds and formulations as defined herein for pharmaceutically active use in treatments that promote endothelial cell-pericyte interactions, particularly vascular stabilization treatments, and / or treatments that promote or maintain pericyte adhesion to blood vessels, as well as to methods for therapeutic treatments that promote endothelial cell-pericyte interactions and vascular stabilization, and / or treatments that promote or maintain pericyte adhesion to blood vessels.

[0050] In one embodiment, the treatment is a preventive therapeutic treatment.

[0051] In one embodiment, the procedure is a therapeutic treatment procedure.

[0052] The vascular stabilizing properties and related medical applicability of the compounds and formulations according to the present invention can be established using an in vivo model with a preclinical mouse model of HHT. By combining electrophysiology and optical microscopy, the ability of pericytes to regulate vasoconstriction in response to electrical stimulation can be quantified. 51 (See also Figures 1 and 3). These experiments may be performed on the whole retina of control mice and heterozygotes or iKO endoglin mice (HHT1 mouse model), as well as on iKO endoglin mice treated with the selected compound (see Figures 3 and 4).

[0053] By comparing the activity of thalidomide and its analogue, lenalidomide, in in vivo models of radiation necrosis and delayed brain injury, we were able to establish the important roles of both carbonyl groups in phthalimide (see Figures 1 and 2). Figure 1 shows that radiation-induced PC dysfunction includes pericyte detachment leading to BBB leakage, and Figure 2 shows that it leads to delayed capillary dilution and neuronal loss.

[0054] Restoring endothelial cell-PC interactions can be used to treat chronic diseases involving vascular changes. The compounds of the present invention can advantageously limit capillary dilution to maintain vascular integrity; limit vascular leakage and inflammation to promote / maintain the blood-brain barrier, particularly the blood-brain barrier (BBB) ​​and the blood-retinal barrier (BRB); reduce the occurrence of vascular hypergrowth and vascular malformations, particularly telangiectasia, low-flow or high-flow vascular malformations; and restore blood flow by limiting tissue hypoperfusion and delayed injury. Therefore, the compounds according to the present invention, or their pharmaceutically acceptable salts or prodrugs, can be used in the pharmaceutical treatment of vascular-related diseases.

[0055] Accordingly, in one embodiment of the present invention, compounds and formulations as defined herein are provided for use as pharmaceuticals in the prophylactic or curative treatment of vascular-related diseases.

[0056] In another embodiment of the present invention, a method for treating a vascular disease in a patient, the method comprising administering to the patient a pharmaceutical compound, or a formulation containing a pharmaceutical compound, wherein the compound is of formula I or II: [ka] [During the ceremony, A 1 , A 2 , A 3 and A 4 Each of them operates independently, CR 1 Or it represents N; A 5 and A 6These are S and CR, which are independent of each other. 1 Or it represents N; R 1 This includes hydrogen, halogens, OH, C1-C8 alkyl, NO2, or NR 2 R 3 This represents, R 2 and R 3 Each of these is independently selected from H, C1-C8 alkyl, C1-C8 alkyl-carbonyl, and C1-C8 alkoxy-carbonyl; R x is H, Y, or CHR 4 R 5 This represents, Y represents a 4-membered, 5-membered, or 6-membered heteroring containing one, two, or three heteroatoms. Y may optionally be substituted with one or more substituents selected from the group consisting of halides, saturated or unsaturated C1-C8 hydrocarbons, C1-C8 alkoxycarbonyls, or C1-C8 alkylcarbonyls, preferably acetyl or propionyl; R 4 This represents H, C1~C8-alkyl, C1~C8-cycloalkyl, C1~C8-cycloalkylalkyl, heteroaryl, C1~C8-heteroaralkyl, C1~C8-heterocycloalkyl, or C1~C8-heterocycloalkylalkyl. R 5 represents H, OH, COOH, C1-C8 alkyl, preferably COOH, i-propyl or t-butyl; optionally, R 4 and R 5 [Except when both are H] A method having the structure shown is provided.

[0057] In a further embodiment of the present invention, a method is provided for promoting, restoring, or maintaining pericyte adhesion to blood vessels in a subject, preferably a method for treating a vascular-related disease, the method comprising administering the compound or formulation defined above to the subject.

[0058] In a further embodiment of the present invention, a method for stabilizing blood vessels in a subject, preferably a method for treating a vascular-related disease, is provided, the method comprising administering a compound or formulation as defined above to the subject.

[0059] Specific examples of vascular diseases include diabetic complications (nephropathy and retinopathy), chronic kidney disease (CKD), cardiomyopathy, central nervous system (CNS) disorders such as stroke, epilepsy, spinal cord injury, dementia, particularly Alzheimer's disease, Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis, radiation necrosis, and small vessel diseases such as autosomal dominant cerebral arteriopathies (Cadasil) and autosomal recessive cerebral arteriopathies (Carasil) with subcortical infarction and leukoencephalopathy, cerebral amyloid angiopathy (CAA), retinal vascular disorders with cerebral leukoencephalopathy and systemic symptoms (RVCL-S), hereditary hemorrhagic telangiectasia (HHT), and cavernous hemangioma (CCM). Most of these conditions still lack the availability of effective treatments suitable for chronic management.

[0060] Detailed explanation of the diagram Figure 1 shows that thalidomide, unlike its derivative lenalidomide, prevents radiotherapy-induced PC detachment and vascular leakage. (A) Diagram of an injection protocol combining vehicle, thalidomide, or lenalidomide with radiotherapy of the brain or eye. (B) Electrically stimulated retinal PC induces local vasoconstriction. The top panel shows representative bright-field images of a single capillary before, during, and after PC electrical stimulation, showing vasoconstriction. The graphs represent the average intensities that induce PC contraction and vasoconstriction in control and irradiated mice treated with vehicle alone, thalidomide, or lenalidomide. (C) Whole brain photographs taken after cadaverine Alexa Fluor-555 circulation, and quantification of tracer accumulation in control and irradiated mice treated with vehicle alone, thalidomide, or lenalidomide 2 hours after tracer circulation. Circulation time was 2 hours. All error bars indicate the mean ± sem (standard error). * P<0.05;** P<0.01; *** P<0.001; **** P<0.0001 indicates a one-way ANOVA and Dunnett's post-hoc test comparing the mean of each group to the control group. ns: No significant difference.

[0061] Figure 2 shows that thalidomide, unlike its derivative lenalidomide, prevents radiotherapy-induced delayed vascular dilution and neuronal loss in the somatosensory cortex (SS) and hippocampal region (Hp). (A) Diagram of the infusion protocol combining vehicle, thalidomide, or lenalidomide with radiotherapy of the brain. (B) Confocal images of brain sections stained for NeuN-positive neurons, showing the somatosensory (SS) cortex and hippocampal region (Hp). (CD) Confocal images of the somatosensory cortex showing GLUT1-stained endothelial cells (C) and NeuN-stained neurons (D) in control and irradiated mice treated with vehicle alone, thalidomide, and lenalidomide. Animals were sacrificed 9 months after receiving a single dose of radiotherapy as shown in A. (E) Quantification of vascular density in the somatosensory cortex. (F) Number of neurons in the cortical region. (GH) Confocal images of the hippocampal region showing endothelial cells stained with GLUT1 (G) and neurons stained with NeuN (H) in control and irradiated mice treated with vehicle alone, thalidomide, and lenalidomide. (I) Quantification of vascular density in the hippocampal region. (F) Number of neurons in the hippocampal region.

[0062] Figure 3 illustrates that thalidomide and selected FDA-approved analogues exhibit different vascular stabilization properties, as illustrated using a mouse model of hereditary hemorrhagic telangiectasia (HHT) type 1. (A) Scheme of treatment plan with tamoxifen (Tx) injection and thalidomide and its derivatives to induce Eng gene deletion (HHT1). (B) Chemical structures of thalidomide (THA), lenalidomide (LENA), pomalidomide (POMA), and apremilast (APRE). (C) Vehicle treated alone or with thalidomide at 4, 40, or 200 mmol / kg-1 Lenalidomide 40, 200, or 400 mmol / kg -1 , pomalidomide 40 mmol.kg -1 Alternatively, Apremilast 40 or 400 mmol / kg -1 Control and Eng-iKO administered e Average intensity for inducing PC contraction in mice. Figure 4 shows (A) selected analogues of phthalimide that restore endothelial cell-pericyte interaction and vascular stability according to the present invention, which are (B) the control and Eng-iKO shown in Figure 3C. e This is as shown by the average intensity that induces PC contraction in mice.

[0063] The methods and procedures described herein may include chronic treatments, such as the chronic administration of pharmaceutical compounds or formulations. They may also include transient or acute treatments. Furthermore, it may be understood that such treatments may be therapeutic and / or prophylactic.

[0064] Another aspect of the present invention relates to the use of organoid and organ-on-a-chip (OoC) models. Organoids and OoCs are in vitro miniaturized organ model systems that have gained considerable interest for disease modeling, personalized medicine, drug testing, and cell therapy. They can be established for an increasingly diverse range of organs, including the brain, kidney, liver, intestine, thyroid, prostate, and airway, from either tissue-resident adult stem cells, biopsy, embryonic stem cells, or induced pluripotent stem cells. Despite considerable success, the limitations on current organoid and OoC systems reaching practical applications remain in their limited maturity and functional levels, and this is also true for vascular networks. The blood vessels of organoids and OoCs appear to be very immature and highly leaky, with a significant reduction in PC coverage.

[0065] As used herein, singular expressions and their similar expressions are intended to include plural expressions unless the context clearly indicates otherwise. The term "and / or" includes any and all combinations of one or more of the items listed relatedly. The term "includes" is understood to identify the presence of the described feature but not to exclude the presence or addition of one or more other features.

[0066] For the purposes of clarity and concise description, features described herein are described as part of the same or distinct embodiments; however, it is understood that the scope of the invention may include embodiments having combinations of all or some of the described features.

[0067] When the absolute stereochemistry of a compound included herein is not specifically indicated, the compound may be a racemic mixture, a mixture of diastereoisomers, or an enantiomerically pure compound. Generally, when the absolute stereochemistry is not specifically indicated, the compound is a racemic mixture. Furthermore, when the relative stereochemistry is not specifically indicated, the compound is a mixture of diastereoisomers.

[0068] The present invention is illustrated by the following non-limiting examples and experiments. [Examples]

[0069] Examples and Experiments Compound Synthesis Exemplary compounds were synthesized by the following general procedure through the condensation of a suitable anhydride (e.g., a suitable phthalic anhydride) with a suitable amine.

[0070] General procedure A for condensation reactions using 3-nitrophthalic anhydride and substituted amines. Equimolar amounts of 3-nitrophthalic anhydride and amine were dissolved in acetic acid and irradiated in a microwave synthesizer at 130°C for 3 hours. The reaction mixture was then concentrated under reduced pressure, added to dichloromethane, and washed with saturated aqueous solution of NaHCO3 and brine, respectively. The organic fraction was dried over MgSO4, and the solvent was evaporated to dryness under reduced pressure. The crude product was finally purified by column chromatography.

[0071] General procedure B for condensation reactions using 3-nitrophthalic anhydride and substituted amines. Equimolar amounts of 3-nitrophthalic anhydride and amine were suspended in THF, and triethylamine (2 equivalents) was added. The suspension was stirred at room temperature until TLC showed complete consumption. The reaction mixture was then concentrated under reduced pressure and added to a 0.6:1 mixture of glacial acetic acid and toluene, and irradiated in a MW synthesizer at 130°C for 0.5 hours. The resulting solution was then concentrated under reduced pressure and subsequently purified by column chromatography.

[0072] General procedure C for nitro reduction using tin(II) chloride dihydrate. The corresponding 3-nitrophthalimide was added to ethyl acetate, and then an excess amount of tin(II) chloride dihydrate was added. The resulting solution was stirred at either 40°C or reflux temperature until complete (TLC). The reaction mixture was then quenched by adding saturated aqueous solution of NaHCO3, the layers were separated, and the aqueous fraction was extracted from ethyl acetate (3x). The combined organic fraction was dried over MgSO4, and the solvent was evaporated to dryness under reduced pressure. Finally, the crude product was purified by column chromatography.

[0073] General procedure D for condensation reactions using ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate and substituted amines. To a THF-stirred solution of ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate (1 equivalent) and triethylamine (2 equivalents), an appropriate amine (1 equivalent) was added all at once. The resulting suspension was stirred overnight at room temperature, after which a clear solution was obtained. The crude reaction mixture was concentrated on silica and purified by column chromatography.

[0074] General procedure E for N-acylation of 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride using various acylclorides. 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride was added to a stirred dichloromethane suspension with triethylamine (2.2 equivalents) and the corresponding asyl chloride (1.2 equivalents). The reaction was stirred at room temperature until a clear solution was obtained and TLC showed that no starting materials remained. The reaction mixture was then concentrated under reduced pressure, and the crude product was purified by column chromatography.

[0075] 2-Isopropyl-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure A under the following conditions: 4-nitroisobenzofuran-1,3-dione (0.10 g, 0.52 mmol) and isopropylamine (0.045 mL, 1 equivalent) were placed in glacial acetic acid (2.5 mL) and irradiated in a microwave synthesizer at 130°C for 3 hours. Yield: 58% (0.07 g, 0.30 mmol), obtained as a colorless solid. 1 H NMR (500MHz, CDCl3) δ 8.12-8.05(m, 2H), 7.91(dd, J=8.1, 7.4Hz, 1H), 4.57(hept, J=7.0Hz, 1H), 1.50(d, J=7.0Hz, 6H).

[0076] 4-amino-2-isopropylisoindoline-1,3-dione(14): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-isopropyl-4-nitroisoindoline-1,3-dione (0.07 g, 0.3 mmol) and tin(II) chloride dihydrate (0.68 g, 10 equivalents) were stirred overnight in ethyl acetate (3 mL) at 40°C. Yield: 84% (0.05 g, 0.25 mmol), obtained as a bright yellow solid. 1H(400MHz, CDCl3) δ 7.37(dd, J=8.3, 7.1Hz, 1H), 7.10(dd, J=7.1, 0.7Hz, 1H), 6.83(dd, J=8.3, 0.7Hz, 1H), 5.06(br s, 2H), 4.46(hept, J=6.9Hz, 1H), 1.45(d, J=7.0Hz, 6H). 13 C NMR (101MHz, CDCl3) δ 170.5, 168.7, 145.0, 135.0, 132.9, 121.1, 112.7, 111.7, 42.6, 20.3. LC-MS(ESI)m / z C 11 H 12 N2O2[M+H] + Calculated value: 204.1, measured value: 205.1. HPLC (Method A): 9.181 mins, purity 99%.

[0077] 2-Isobutyl-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure A under the following conditions: 4-nitroisobenzofuran-1,3-dione (0.15 g, 0.78 mmol) and isobutylamine (0.068 g, 1.2 equivalents) were placed in glacial acetic acid (2 mL) and irradiated at 130°C for 3 hours in a microwave synthesizer. Yield: 89% (0.17 g, 0.69 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.15-8.13(m, 1H), 8.15-8.09(m, 1H), 7.94(dd, J=8.2, 7.4Hz, 1H), 3.55(d, J=7.4Hz, 2H), 2.24-2.05(m, 1H), 0.96(d, J=6.8Hz, 6H).

[0078] 4-amino-2-isobutylisoindoline-1,3-dione(15): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-Isobutyl-4-nitroisoindoline-1,3-dione (0.171 g, 0.69 mmol) and tin(II) chloride dihydrate (1.6 g, 10 equivalents) were stirred in ethyl acetate (3 mL) at 40°C for 72 hours. Yield: 87% (0.13 g, 0.60 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, CDCl3) δ 7.39(dd, J=8.3, 7.1Hz, 1H), 7.13(dd, J=7.1, 0.7Hz, 1H), 6.85(dd, J=8.3, 0.7Hz, 1H), 5 .02(s, 2H), 3.43(d, J=7.4Hz, 2H), 2.08(dh, J=13.7, 6.9Hz, 1H), 0.92(d, J=6.7Hz, 6H). 13 C NMR (101MHz, CDCl3) δ 170.62, 169.01, 145.07, 135.16, 132.86, 121.15, 112.87, 111.57, 45.04, 27.99, 20.25. LC-MS(ESI)m / z C 12 H 14 N2O2[M+H] + Calculated value: 218.1, measured value: 219.0. HPLC (Method A): 9.737 mins, purity 97%.

[0079] 4-Nitro-2-phenylisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure A under the following conditions: 4-nitroisobenzofuran-1,3-dione (0.15 g, 0.78 mmol) and aniline (0.045 mL, 1 equivalent) were placed in glacial acetic acid (2.5 mL) and irradiated in a microwave synthesizer at 130°C for 3 hours. Yield: 75% (0.16 g, 0.58 mmol), obtained as a colorless crystalline solid. 1 H NMR (400MHz, CDCl3) δ 8.22 (dd, J=7.5, 1.0Hz, 1H), 8.16 (dd, J=8.2, 0.9Hz, 1H), 7.99 (dd, J=8.1, 7.5Hz, 1H), 7.57-7.48 (m, 2H), 7.47-7.40 (m, 3H).

[0080] 4-amino-2-phenylisoindoline-1,3-dione(16): [ka] The compound was synthesized according to general procedure C under the following conditions: 4-nitro-2-phenylisoindoline-1,3-dione (0.15 g, 0.56 mmol) and tin(II) chloride dihydrate (0.51 g, 4 equivalents) were stirred in ethyl acetate (15 mL) at 40°C for 16 hours. Yield: 79% (0.11 g, 0.44 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, CDCl3) δ 7.42-7.26 (m, 6H), 7.14 (dd, J=7.1, 0.7Hz, 1H), 6.80 (dd, J=8.3, 0.7Hz, 1H), 5.13 (br s, 2H). 13 C NMR (101MHz, CDCl3) δ 169.26, 167.61, 145.84, 135.66, 132.51, 131.93, 129.19, 127.97, 126.68, 121.39, 113.23, 111.00. LC-MS(ESI)m / z C 14 H 10 N2O2[M+H] + Calculated value: 238.1, measured value: 239.0. HPLC (Method A): 9.41 min, purity 98%.

[0081] tert-butyl(S)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)pyrrolidine-1-carboxylate: [ka] The compound was synthesized according to general procedure B under the following conditions: tert-butyl(S)-3-aminopyrrolidine-1-carboxylate (0.50 g, 2.68 mmol), 4-nitroisobenzofuran-1,3-dione (0.57 g, 1.1 equivalents), and triethylamine (0.7 mL, 2 equivalents). Yield: 93% (0.90 g, 2.50 mmol), obtained as a light brown oily substance. 1H NMR (400MHz, CDCl3) δ 8.22-8.08(m, 2H), 7.96(t, J=7.7Hz, 1H), 4.89(p, J=8.2Hz, 1H), 3.78-3.64(m, 3H), 3. 49-3.37(m, 1H), 2.61(p, J=9.1Hz, 1H), 2.16(d, J=22.8Hz, 1H), 1.47(d, J=7.3Hz, 9H).

[0082] (S)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)pyrrolidine-1-ium chloride: [ka] tert-butyl(S)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)pyrrolidine-1-carboxylate (0.90 g, 2.49 mmol) was added to methanolic HCl (4N, 25 mL) and stirred at room temperature for 1 hour. After 1 hour of stirring, the reaction mixture was filtered, the solid was washed with ELISA, and then dried under reduced pressure to obtain the title compound as a white solid. Yield: 81% (0.60 g, 2.02 mmol). 1 H NMR (400MHz, DMSO-D6) δ 9.73(br s, 1H), 9.31(br s, 1H), 8.30(dd, J=8.1, 0.8Hz, 1H), 8.18(dd, J=7.5, 0.8Hz, 1H), 8.08(dd, J=8.0, 7.6 Hz, 1H), 4.97-4.85(m, 1H), 3.56-3.41(m, 3H), 3.35-3.25(m, 1H), 2.38-2.20(m, 2H).

[0083] (S)-2-(1-acetylpyrrolidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] (S)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)pyrrolidine-1-ium chloride (0.15 g, 0.50 mmol) was added to a stirred suspension in DCM (10 mL) with triethylamine (0.05 mL, 2 equivalents) and acetyl chloride (0.14 mL, 1.4 equivalents). The resulting solution was stirred for 2 hours. The reaction mixture was then concentrated under reduced pressure to obtain the crude title compound as a crystalline white solid. Yield: 0.20 g, 0.50 mmol, quantitative. 1 H NMR (400MHz, CDCl3) δ 8.18-8.09(m, 2H), 7.96(q, J=7.6Hz, 1H), 4.95(dp, J=15.6, 8.1Hz, 1H), 4.02-3.72(m, 3H), 3.68-3.42(m, 1H), 2.81-2.52(m, 1H), 2.43-2.28(m, 1H), 2.09(d, J=14.0Hz, 3H).

[0084] (S)-2-(1-acetylpyrrolidine-3-yl)-4-aminoisoindoline-1,3-dione(25): [ka] The compound was synthesized according to general procedure C under the following conditions: (S)-2-(1-acetylpyrrolidine-3-yl)-4-nitroisoindorin-1,3-dione (0.15 g, 0.50 mmol) and tin(II) chloride dihydrate (0.45 g, 4 equivalents) were stirred under reflux in ethyl acetate (60 mL) for 6 hours. The crude product was purified by column chromatography and eluted using 3% MeOH in DCM as the eluent to obtain the title compound as a yellow film. Yield: 63% (0.13 g, 0.47 mmol), obtained as a bright yellow solid. 1H NMR (400MHz, DMSO-D6) δ 7.42 (ddd, J=8.5, 7.0, 2.4Hz, 1H), 7.00-6.91 (m, 2H), 6.48 (d, J=5.1Hz, 2H), 4.73 (dp, J=27.4, 8.0Hz, 1H), 3.80-3.54 (m, 3H), 3.50 (dt, J= 9.9, 7.7Hz, 0.5H), 3.30 (dt, J=11.6, 8.1Hz, 0.5H), 2.47-2.27 (m, 1H), 2.13 (ddtd, J=32.1, 12.1, 7.6, 4.4Hz, 1H), 1.94 (d, J=14.1Hz, 3H). 13 C NMR (101MHz, DMSO) δ 169.29, 168.15, 167.96, 167.89, 167.85, 146.55, 146.52, 135.20, 135.16, 132.19, 132.15, 121.43, 121.3 9. 110.67, 110.65, 108.82, 108.75, 48.56, 48.09, 47.37, 46.79, 45.70, 43.96, 29.05, 27.44, 22.32, 21.88. 1 H NMR (500MHz, DMSO-D6, 100℃) δ 7.43 (dd, J=8.4, 7.0Hz, 1H), 7.01 (d, J=8.4Hz, 1H), 6.97 (d, J=7.0Hz, 1H), 6.24 (s, 2H), 4.85-4.62 (m, 1H ), 4.02-3.59 (m, 3H), 3.53 (s, 0.5H), 3.36 (s, 0.5H), 2.48-2.32 (m, 1H), 2.30-2.05 (m, 1H), 1.96 (s, 3H). LC-MS(ESI)m / z C 14 H 15 N3O3[M+H] + The calculated value is 273.1 and the measured value is 274.0. HPLC (method A): 6.92 points, purity 99%.

[0085] tert-ブチル(R)-3-(4-ニトロ-1,3-ジオキソイソインドリン-2-イル)ピロリジン-1-カルボキシレート:

change

[0086] (R)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)pyrrolidine-1-ium chloride: [ka] tert-butyl(R)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)pyrrolidine-1-carboxylate (0.80 g, 2.21 mmol) was added to methanolic HCl (4N, 25 mL) and stirred at room temperature for 1 hour. After stirring for 1 hour, the reaction mixture was filtered, the solid was washed with ethyl acetate, and then dried under reduced pressure to obtain the title compound as a white solid. Yield: 84% (0.55 g, 1.85 mmol). 1 H NMR (400MHz, DMSO-D6) δ 9.64(s, 1H), 9.24(s, 1H), 8.30(dd, J=8.1, 0.8Hz, 1H), 8.18(dd, J=7.5, 0.9Hz, 1H), 8.08(dd, J=8.0 , 7.6Hz, 1H), 4.92(tt, J=8.5, 6.0Hz, 1H), 3.60-3.40(m, 3H), 3.34-3.25(m, 1H), 2.40-2.19(m, 2H).

[0087] (R)-2-(1-acetylpyrrolidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] (R)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)pyrrolidine-1-ium chloride (0.15 g, 0.50 mmol) was added to a stirred suspension in DCM (10 mL) with triethylamine (0.05 mL, 2 equivalents) and acetyl chloride (0.14 mL, 1.4 equivalents). The resulting solution was stirred for 2 hours. The reaction mixture was then concentrated under reduced pressure to obtain the crude title compound as a crystalline white solid. Yield: 0.20 g, 0.50 mmol, quant. 1 H NMR (400MHz, CDCl3) δ 8.13(ddd, J=7.8, 6.3, 1.7Hz, 2H), 8.01-7.91(m, 1H), 4.95(dp, J=15.6, 8.2Hz, 1H), 4.02-3. 69(m, 3H), 3.66-3.44(m, 1H), 2.74-2.54(m, 1H), 2.40-2.28(m, 1H), 2.09(d, J=14.1Hz, 3H).

[0088] (R)-2-(1-acetylpyrrolidine-3-yl)-4-aminoisoindoline-1,3-dione(27): [ka] The compound was synthesized according to general procedure C under the following conditions: (R)-2-(1-acetylpyrrolidine-3-yl)-4-nitroisoindoline-1,3-dione (0.15 g, 0.50 mmol) and tin(II) chloride dihydrate (0.45 g, 4 equivalents) were stirred in ethyl acetate (60 mL) under reflux for 16 hours. The crude product was purified by column chromatography and eluted using 3% MeOH in DCM as the eluent to obtain the title compound as a yellow film. Yield: 99% (0.13 g, 0.49 mmol), obtained as a bright yellow solid. 1H NMR (400MHz, DMSO-D6) δ 7.42 (ddd, J=8.3, 7.0, 2.5Hz, 1H), 7.00-6.91 (m, 2H), 6.48 (d, J=5.1Hz, 2H), 4.73 (dp, J=27.4, 8.0Hz, 1H), 3.82-3.54 (m, 3H), 3.50 (dt, J=9.9, 7.8Hz, 0.5H), 3.30 (dt, J=11.6, 8.1Hz, 0.5H), 2.48-2.27 (m, 1H), 2.24-1.99 (m, 1H), 1.94 (d, J=14.1Hz, 3H). 13 C NMR (101MHz, DMSO-D6) δ 169.29, 168.14, 167.96, 167.85, 146.55, 146.52, 135.20, 135.16, 132.19, 132.14, 121.44, 121.39, 11 0.67, 110.65, 108.80, 108.75, 48.56, 48.09, 47.36, 46.79, 45.70, 43.96, 29.05, 27.44, 22.32, 21.88. 1 H NMR (500MHz, DMSO-D6, 100℃) δ 7.43 (dd, J=8.4, 7.0Hz, 1H), 7.01 (d, J=8.4Hz, 1H), 6.97 (d, J=7.1Hz, 1H), 6.25 (br s, 2H), 4.85-4.61 (m, 1H), 3.99-3.59 (m, 3H), 3.59-3.44 (m, 0.5H), 3.44-3.25 (m, 0.5H), 2.47-2.31 (m, 1H), 2.29-2.09 (m, 1H), 1.96 (br s, 3H). LC-MS(ESI)m / z C 14 H 15 N3O3[M+H] + The calculated value is 273.1 and the measured value is 274.0. HPLC (method A): 7.18 points, purity 98%.

[0089] tert-ブチル4-(4-ニトロ-1,3-ジオキソイソインドリン-2-イル)ピペリジン-1-カルボキシレート:

change

[0090] 4-(4-nitro-1,3-dioxoisoindorin-2-yl)piperidine-1-ium chloride: [ka] tert-butyl 4-(4-nitro-1,3-dioxoisoindolin-2-yl)piperidine-1-carboxylate (0.50 g, 1.33 mmol) was added to methanolic HCl (4N, 20 mL) and stirred at room temperature for 1 hour. After stirring for 1 hour, the reaction mixture was filtered, the solid was washed with ELISA, and then dried under reduced pressure to obtain the title compound as a white solid. Yield: 56% (0.23 g, 0.75 mmol). 1 H NMR (400MHz, DMSO-D6) δ 9.17(s, 1H), 8.59(s, 1H), 8.27(dd, J=8.1, 0.8Hz, 1H), 8.16(dd, J=7.5, 0.9Hz, 1H), 8.10-8.01(m, 1H), 4.34(tt , J=12.1, 3.7Hz, 1H), 3.36(d, J=12.5Hz, 2H), 3.04(q, J=12.4Hz, 2H), 2.48-2.35(m, 2H), 1.91(d, J=12.8Hz, 2H).

[0091] 2-(1-acetylpiperidine-4-yl)-4-nitroisoindoline-1,3-dione: [ka] To a stirred suspension of 4-(4-nitro-1,3-dioxoisoindorin-2-yl)piperidine-1-ium chloride (0.15 g, 0.48 mmol) in DCM (20 mL), triethylamine (0.16 mL, 2.4 equivalents) and acetyl chloride (0.08 mL, 2.3 equivalents) were added, and the solution was stirred for 1 hour. The reaction mixture was then washed with brine (2x), the organic fraction was dried over MgSO4, and evaporated to dryness under reduced pressure to obtain the title compound as a white crystalline solid. Yield: 99% (0.15 g, 0.48 mmol). 1 H NMR (400MHz, CDCl3) δ 8.14-8.07(m, 2H), 7.96-7.88(m, 1H), 4.84(ddd, J=11.1, 4.4, 2.2Hz, 1H), 4.38(tt, J=12.2, 4.1Hz, 1H), 4.02-3.92(m , 1H), 3.16(td, J=13.4, 2.6Hz, 1H), 2.60(td, J=13.2, 2.6Hz, 1H), 2.52-2.28(m, 2H), 2.15(s, 3H), 1.85-1.71(m, 2H).

[0092] 2-(1-acetylpiperidine-4-yl)-4-aminoisoindoline-1,3-dione(29): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-acetylpiperidine-4-yl)-4-nitroisoindorin-1,3-dione (0.15 g, 0.48 mmol) and tin(II) chloride dihydrate (0.43 g, 4 equivalents) were stirred under reflux in ethyl acetate (30 mL) for 6 hours. The crude product was purified by column chromatography, and the title compound was obtained as a yellow solid by elution using a gradient of 0-2% MeOH in DCM as the eluent. Yield: 99% (0.14 g, 0.47 mmol), obtained as a bright yellow solid. 1H NMR (400MHz, CDCl3) δ 7.41(dd, J=8.3, 7.2Hz, 1H), 7.12(dd, J=7.1, 0.5Hz, 1H), 6.85(dd, J=8.3, 0.5Hz, 1H), 5.26(s, 2H), 4.81(ddd, J=11.1, 4.4, 2.2Hz, 1H), 4.27(tt, J=12.2, 4.1Hz, 1H), 3.95(dd d, J=11.4, 4.4, 2.1Hz, 1H), 3.14(td, J=13.4, 2.6Hz, 1H), 2.59(td, J=13.2, 2.6Hz, 1H) , 2.39(dqd, J=39.9, 12.6, 4.4Hz, 2H), 2.14(s, 3H), 1.75(tdd, J=11.7, 4.2, 2.3Hz, 2H). 13 C NMR (101MHz, CDCl3) δ 170.16, 168.97, 168.42, 145.41, 135.35, 132.54, 121.19, 112.81, 111.11, 48.44, 46.23, 41.40, 29.58, 28.98, 21.63. LC-MS(ESI)m / z C 15 H 17 N3O3[M+H] + Calculated value: 287.1, measured value: 288.0. HPLC (Method A): 7.68 mins, purity 99%.

[0093] tert-butyl 3-methyl-3-(4-nitro-1,3-dioxoisoindolin-2-yl)azetidine-1-carboxylate: [ka] The compound was synthesized according to general procedure B under the following conditions: tert-butyl 3-amino-3-methylazetidine-1-carboxylate (0.50 g, 2.68 mmol), 4-nitroisobenzofuran-1,3-dione (0.52 g, 1.0 equivalent), and triethylamine (0.75 mL, 2 equivalents). Yield: 79% (1.16 g, 3.09 mmol), obtained as a white solid. 11H NMR (400 MHz, DMSO-D6) δ 8.26 (dd, J = 8.0, 0.9 Hz, 1H), 8.13 (dd, J = 7.4, 0.9 Hz, 1H), 8.04 (dd, J = 8.0, 7.5 Hz, 1H), 4.37 (d, J = 8.7 Hz, 2H), 3.82 (d, J = 8.7 Hz, 2H), 1.59 (s, 3H), 1.38 (s, 9H).

[0094] 3-Methyl-3-(4-nitro-1,3-dioxoisoindolin-2-yl)azetidin-1-ium chloride:

Chemical formula

[0095] 2-(1-Acetyl-3-methylazetidin-3-yl)-4-nitroisoindoline-1,3-dione:

Chemical formula

[0096] 2-(1-acetyl-3-methylazetidine-3-yl)-4-aminoisoindoline-1,3-dione(33): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-acetyl-3-methylazetidine-3-yl)-4-nitroisoindoline-1,3-dione (0.26 g, 0.84 mmol) and tin(II) chloride dihydrate (0.76 g, 4 equivalents) were stirred under reflux in ethyl acetate (25 mL) for 16 hours. The crude product was purified by column chromatography and eluted with 100% ethyl acetate as the eluent to obtain the title compound as a bright yellow solid. Yield: 95% (0.22 g, 0.80 mmol). 1H NMR (400MHz, CDCl3) δ 7.41(dd, J=8.3, 7.1Hz, 1H), 7.09(dd, J=7.1, 0.7Hz, 1H), 6.89(dd, J=8.4, 0.8Hz, 1H), 5.41(br s, 2H), 4.71(d, J=9.3Hz, 1H), 4.53(d, J=10.6Hz, 1H), 4.21(d, J=9.3Hz, 1H), 4.15(d, J=10.6Hz, 1H), 1.91(s, 3H), 1.70(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.98, 169.45, 167.82, 145.70, 135.46, 132.61, 121.44, 112.60, 110.84, 60.80, 58.46, 49.50, 24.73, 19.02. LC-MS(ESI)m / z C 14 H 15 N3O3[M+H] + Calculated value: 273.1, measured value: 274.0. HPLC (Method A): 7.34 mins, purity 99%.

[0097] Ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate: [ka] To a 17 mL DCM-stirred suspension of 3-nitrophthalimide (2.0 g, 10.4 mmol), triethylamine (6.5 mL, 4.5 equivalents) and DMAP (0.13 g, 0.1 equivalents) were added. The mixture was then cooled to 0°C using an ice bath, and ethyl chloroformate (5 mL, 5 equivalents) was added dropwise. After the addition was complete, the solution was maintained at 0°C for 30 minutes, and the reaction mixture was then diluted with ethyl acetate (250 mL), followed by washing with HCl (aqueous solution, 3% v / v) and brine. The organic fraction was then dried over MgSO4 and concentrated under reduced pressure. The crude product was purified by recrystallization from an HCl / hexane mixture. Yield: 70% (1.91 g, 7.23 mmol), obtained as a light brown solid. 1H NMR (400MHz, CDCl3) δ 8.24 (dd, J=7.6, 0.9Hz, 1H), 8.19 (dd, J=8.0, 1.0Hz, 1H), 8.03 (t, J=7.8Hz, 1H), 4.51 (q, J=7.1Hz, 2H), 1.45 (t, J=7.1Hz, 3H).

[0098] Ethyl 1,3-dioxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate: [ka] To a stirred suspension of 1H-pyrrolo[3,4-c]pyridine-1,3(2H)-dione (2.5 g, 16.9 mmol) and triethylamine (3.1 mL, 1.3 equivalents) in DMF (9 mL), ethyl chloroformate (2.0 mL, 1.2 equivalents) was added dropwise at 0°C. After the addition was complete, the solution was maintained at 0°C for 30 minutes, and then the reaction mixture was maintained at room temperature for 1 hour. The reaction mixture was then poured onto ice and extracted with ethyl acetate (3 x 200 mL), and the combined organic fraction was washed with brine. The organic fraction was dried over MgSO4 and concentrated under reduced pressure. The crude product was purified by column chromatography and eluted with 40 / 60 ethyl acetate in pentane to obtain the title compound as a yellow crystalline solid. Yield: 30% (1.11 g, 5.04 mmol). 1 H NMR (400MHz, CDCl3) δ 9.30(s, 1H), 9.17(d, J=4.9Hz, 1H), 7.88(d, J=4.8Hz, 1H), 4.52(q, J=7.1Hz, 2H), 1.46(t, J=7.1Hz, 3H).

[0099] 2-(1-acetylazetidine-3-yl)-1H-pyrrolo[3,4-c]pyridine-1,3(2H)-dione(4): [ka] The compound was synthesized according to general procedure D using the following reagents: ethyl 1,3-dioxo-1,3-dihydro-2H-pyrrolo[3,4-c]pyridine-2-carboxylate (0.15 g, 0.68 mmol), triethylamine (0.19 mL, 2 equivalents), and 1-acetylazetidine-3-aminium chloride (0.1 g, 1 equivalent). The reaction mixture was stirred at room temperature for 2 hours, and then heated at 50°C for 2 hours. Yield: 30% (0.05 g, 0.21 mmol), obtained as a grayish-white solid. 1 H NMR (400MHz, CDCl3δ 9.15(d, J=1.1Hz, 1H), 9.08(d, J=4.9Hz, 1H), 7.76(dd, J=4.8, 1.2Hz, 1H), 5.09(tt, J=8.7, 6.2Hz, 1H), 4.70(d d, J=8.7, 6.1Hz, 1H), 4.49(dd, J=10.1, 6.3Hz, 1H), 4.42(t, J=8.7Hz, 1H), 4.32(t, J=9.4Hz, 1H), 1.92(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.62, 166.56, 166.26, 156.15, 145.15, 138.97, 125.40, 117.09, 54.53, 52.63, 38.79, 19.06. LC-MS(ESI)m / z C 12 H 11 N3O3[M+H] + Calculated value: 245.1, measured value: 246.1. HPLC (Method B): 7.78 mins, purity 96%.

[0100] (S)-4-methyl-2-(4-nitro-1,3-dioxoisoindolin-2-yl)pentanoic acid: [ka] The compound was synthesized according to general procedure D using the following reagents: ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate (0.4 g, 1.5 mmol), triethylamine (0.42 mL, 2 equivalents), and L-leucine (0.20 g, 1 equivalent). After stirring overnight at room temperature, the reaction mixture was heated to 130°C for 15 minutes. Yield: 52% (0.24 g, 0.78 mmol), obtained as a colorless oil. 1 H NMR (400MHz, CDCl3) δ 8.17-8.11(m, 2H), 7.94(t, J=7.8Hz, 1H), 5.00(ddd, J=11.5, 4.4, 1.4Hz, 1H), 1.97(dddd , J=14.5, 10.3, 4.4, 1.2Hz, 1H), 1.55-1.43(m, 1H), 1.34-1.20(m, 1H), 1.02-0.87(m, 6H).

[0101] (S)-2-(4-amino-1,3-dioxoisoindolin-2-yl)-4-methylpentanoic acid (17): [ka] The compound was synthesized according to general procedure C under the following conditions: (S)-4-methyl-2-(4-nitro-1,3-dioxoisoindorin-2-yl)pentanoic acid (0.23 g, 0.75 mmol) and tin(II) chloride dihydrate (0.68 g, 4 equivalents) were stirred in ethyl acetate (10 mL) at 40°C for 16 hours. The product was purified by flash column chromatography using a C18 cartridge and eluted using a 10-90% CH3CN gradient in H2O. Yield: 63% (0.13 g, 0.47 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, CDCl3) δ 7.40(t, J=7.7Hz, 1H), 7.15(d, J=7.1Hz, 1H), 6.87(d, J=8.3Hz, 1H), 4.92(dd, J=11.5, 4.3Hz, 1H), 2.31(dd d, J=15.1, 11.6, 4.1Hz, 1H), 1.90(ddd, J=14.4, 10.2, 4.4Hz, 1H), 1.57-1.40(m, 1H), 0.92(t, J=6.8Hz, 6H).13 13C NMR (101 MHz, CDCl3) δ 175.88, 169.50, 168.03, 145.10, 135.55, 132.46, 121.68, 113.50, 111.34, 50.15, 37.20, 25.14, 23.26, 21.10. LC-MS (ESI) m / z C 14 H 16 N2O4 [M + H] + Calculated value 276.1, measured value 277.0. HPLC (Method A): 9.43 minutes, purity 97%.

[0102] (S)-4,4-Dimethyl-2-(4-nitro-1,3-dioxoisoindolin-2-yl)pentanoic acid:

Chemical Structure

[0103] (S)-2-(4-Amino-1,3-dioxoisoindolin-2-yl)-4,4-dimethylpentanoic acid (30):

Chemical Structure

[0104] (S)-3-Cyclopropyl-2-(4-Nitro-1,3-Dioxoisoindolin-2-yl)propanoic acid: [ka] The compound was synthesized according to general procedure D using the following reagents: ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate (1.02 g, 3.87 mmol), triethylamine (1.08 mL, 2 equivalents), and (S)-2-amino-3-cyclopropylpropanoic acid (0.50 g, 1 equivalent). After stirring at room temperature under an N2 atmosphere for 2 hours, the reaction mixture was irradiated in an MW synthesizer at 130°C for 5 minutes. Yield: 25% (0.30 g, 1.0 mmol), obtained as a colorless oil. 1 H NMR (400MHz, CDCl3) δ 11.11-10.84(m, 1H), 8.18(dd, J=3.2, 0.9Hz, 1H), 8.17-8.13(m, 1H), 8.02-7.93(m, 1H), 5.05(dd, J=10.3, 5.4Hz, 1H), 2.24-2.12(m, 2H), 0.73-0.57(m, 1H), 0.49-0.32(m, 2H), 0.12(td, J=9.1, 8.7, 4.9Hz, 1H), 0.08-0.00(m, 1H).

[0105] (S)-2-(4-amino-1,3-dioxoisoindolin-2-yl)-3-cyclopropylpropanoic acid (31): [ka] The compound was synthesized according to general procedure C under the following conditions: (S)-3-cyclopropyl-2-(4-nitro-1,3-dioxoisoindorin-2-yl)propanoic acid (0.30 g, 1.0 mmol) and tin(II) chloride dihydrate (0.89 g, 4 equivalents) were stirred under reflux in ethyl acetate (40 mL) for 6 hours. The crude product was purified by column chromatography and eluted with a 2% MeOH and 0.5% AcOH dichloromethane solution to obtain the title compound as a yellow solid. Yield: 89% (0.24 g, 0.88 mmol). 1H(400MHz, CDCl3) δ 7.40(dd, J=8.3, 7.1Hz, 1H), 7.15(dd, J=7.2, 0.7Hz, 1H), 6.86(dd, J=8.3, 0.7Hz, 1H), 4.96(dd, J=10.8, 4.8Hz, 1H), 2.17(d dd. 13 C NMR (101MHz, CDCl3) δ 175.43, 169.56, 168.07, 145.63, 135.51, 132.45, 121.44, 113.18, 110.97, 52.03, 33.45, 8.25, 4.76, 3.64. LC-MS(ESI)m / z C 14 H 14 N2O4[M+H] + Calculated value: 275.1, measured value: 275.0. HPLC (Method A): 8.81 mins, purity 99%.

[0106] (S)-2-(4-nitro-1,3-dioxoisoindolin-2-yl)-3-(thiophen-2-yl)propanoic acid: [ka] The compound was synthesized according to general procedure D using the following reagents: ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate (0.77 g, 2.92 mmol), triethylamine (0.81 mL, 2 equivalents), and (S)-2-amino-3-(thiophen-2-yl)propanoic acid (0.50 g, 1 equivalent). After stirring at room temperature under an N2 atmosphere for 2 hours, the reaction mixture was irradiated in a MW synthesizer at 130°C for 5 minutes. Yield: 78% (0.79 g, 2.28 mmol), obtained as a light brown solid. 1H NMR (400MHz, CDCl3) δ 10.51(s, 1H), 8.14(dd, J=8.1, 1.0Hz, 1H), 8.10(dd, J=7.5, 0.9Hz, 1H), 7.92(dd, J=8.1, 7.5Hz, 1H ), 7.09(dd, J=4.7, 1.7Hz, 1H), 6.87-6.80(m, 2H), 5.22(dd, J=11.5, 4.8Hz, 1H), 3.97-3.71(m, 2H).

[0107] (S)-2-(4-amino-1,3-dioxoisoindolin-2-yl)-3-(thiophen-2-yl)propanoic acid (32): [ka] The compound was synthesized according to general procedure C under the following conditions: (S)-2-(4-nitro-1,3-dioxoisoindolin-2-yl)-3-(thiophen-2-yl)propanoic acid (0.35 g, 1.0 mmol) and tin(II) chloride dihydrate (0.92 g, 4 equivalents) were stirred under reflux in ethyl acetate (40 mL) for 6 hours. The crude product was purified by column chromatography and eluted using a 2-10% MeOH gradient in dichloromethane. The resulting solid was then washed with heated dichloromethane to obtain the title compound as a yellow solid. Yield: 46% (0.15 g, 0.47 mmol). 1 H NMR (400MHz, DMSO-D6) δ 13.36(br s, 1H), 7.43(dd, J=8.5, 7.0Hz, 1H), 7.26(dd, J=5.0, 1.4Hz, 1H), 6.96(dd, J=14.9, 7.7Hz, 2H), 6.90-6.82(m, 2H), 6.51(br s, 2H), 4.92(dd, J=9.7, 6.4Hz, 1H), 3.70-3.55(m, 2H). 13 C NMR (101MHz, DMSO-D6) δ 170.00, 168.65, 167.46, 146.73, 139.26, 135.51, 131.81, 126.94, 126.19, 124.93, 121.79, 110.98, 108.25, 52.57, 28.26. LC-MS(ESI)m / z C 15 H 12N2O4S [M+H] + Calculated value: 316.1, measured value: 317.0. HPLC (Method A): 9.11 mins, purity 99%.

[0108] tert-butyl 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-carboxylate: [ka] The compound was synthesized according to general procedure D using the following reagents: ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate (0.1 g, 0.38 mmol), triethylamine (0.11 mL, 2 equivalents), and tert-butyl 3-aminoazetidine-1-carboxylate hydrochloride (0.07 g, 1 equivalent). The product was purified by column chromatography using a gradient of 0-1% MeOH in dichloromethane. Yield: 82% (0.11 g, 0.31 mmol), obtained as a white solid. 1 H NMR (400MHz, CDCl3) δ 8.18-8.15(m, 1H), 8.15-8.13(m, 1H), 7.96(dd, J=8.2, 7.4Hz, 1H), 5.05(tt, J=8 .5, 6.2Hz, 1H), 4.51(dd, J=9.0, 6.3Hz, 2H), 4.26(t, J=8.9Hz, 2H), 1.48(s, 9H).

[0109] tert-butyl 3-(4-amino-1,3-dioxoisoindorin-2-yl)azetidine-1-carboxylate (21): [ka] The compound was synthesized according to general procedure C under the following conditions: tert-butyl 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-carboxylate (0.09 g, 0.27 mmol) and tin(II) chloride dihydrate (0.37 g, 6 equivalents) were stirred in ethyl acetate (15 mL) at 40°C for 3 hours. The product was purified by flash column chromatography using a C18 cartridge and eluted using a gradient of 10-90% CH3CN in H2O. Yield: 40% (0.03 g, 0.11 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, CDCl3) δ 7.41(dd, J=8.2, 7.3Hz, 1H), 7.12(d, J=7.1Hz, 1H), 6.86(d, J=8.4Hz, 1H), 4.96(tt, J=8.6, 6.2Hz, 1H), 4.50(dd, J=8.9, 6.2Hz, 2H), 4.20(t, J=8.7Hz, 2H), 1.47(s, 9H). 13 C NMR (101MHz, CDCl3) δ 169.76, 168.10, 156.58, 145.65, 135.58, 132.40, 121.45, 112.97, 110.86, 79.92, 54.11, 38.30, 28.51. LC-MS(ESI)m / z C 16 H 19 N3O4[M+H] + Calculated value: 317.1, Measured value: 218.0 [M-Boc+H] + . HPLC (Method A): 9.92 min, 94% purity.

[0110] 4-amino-2-(azetidine-3-yl)isoindoline-1,3-dione hydrochloride (34): [ka] tert-butyl 3-(4-amino-1,3-dioxoisoindolin-2-yl)azetidine-1-carboxylate (0.10 g, 0.32 mmol) was stirred for 2 hours in a 4N HCl solution with MeOH (20 mL). The resulting suspension was concentrated under reduced pressure and co-evaporated with ELISA (2x) to obtain the title compound as the HCl salt. Yield: 88% (0.08 g, 0.28 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, DMSO-D6) δ 9.63(br s, 1H), 9.18(br s, 1H), 7.45(dd, J=8.5, 7.0Hz, 1H), 7.02(dd, J=8.5, 0.7Hz, 1H), 6.98(dd, J=7.0, 0.7Hz, 1H), 6.36(br s, 3H), 5.01(p, J=8.3Hz, 1H), 4.59-4.46(m, 2H), 4.25-4.08(m, 2H). 13 C NMR (101MHz, DMSO-D6) δ 168.90, 167.72, 146.78, 135.42, 132.09, 121.68, 110.92, 108.65, 49.84, 39.99. LC-MS(ESI)m / z C 11 H 11 N3O2[M+H] + Calculated value: 217.1, Measured value: 218.0 [M+H] + . HPLC (Method B): 1.19 min, 95% purity.

[0111] tert-butyl(S)-3-(4-nitro-1,3-dioxoisoindoline-2-yl)piperidine-1-carboxylate: [ka] The compound was synthesized according to general procedure D using the following reagents: ethyl 4-nitro-1,3-dioxoisoindoline-2-carboxylate (0.66 g, 2.50 mmol), triethylamine (0.70 mL, 2 equivalents), and tert-butyl(S)-3-aminopiperidine-1-carboxylate (0.50 g, 1 equivalent). Yield: 99% (0.93 g, 2.48 mmol), obtained as a colorless oil. 1H NMR (400MHz, CDCl3) δ 8.17-8.08(m, 2H), 7.94(dd, J=8.2, 7.4Hz, 1H), 4.88-4.68(m, 1H), 4.22(tt, J=11.9, 4.3Hz, 1H), 3.65-3.30(m, 1 H), 2.88-2.55(m, 1H), 2.32(tt, J=12.5, 6.3Hz, 1H), 1.93-1.78(m, 2H), 1.68-1.56(m, 1H), 1.46(d, J=1.4Hz, 9H).

[0112] (S)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)piperidine-1-ium chloride: [ka] To a stirred solution of tert-butyl(S)-3-(4-nitro-1,3-dioxoisoindorin-2-yl)piperidine-1-carboxylate (0.90 g, 2.40 mmol) in methanol (10 mL), 4N HCl in dioxane (10 mL) was carefully added, and the mixture was stirred for 1 hour. The reaction mixture was then concentrated under reduced pressure to obtain the title compound as a white solid. Yield: 75% (0.56 g, 1.81 mmol). 1 H NMR (400MHz, DMSO-D6) δ 9.57 (br s, 1H), 9.42 (br s, 1H), 8.29(dd, J=8.0, 0.9Hz, 1H), 8.17(dd, J=7.5, 0.9Hz, 1H), 8.09-8.04(m, 1H), 4.47(ddt, J=16.2, 10.1, 3.7Hz, 1H), 3.34(br s, 2H), 3.25 (d, J=12.5Hz, 1H), 2.83 (t, J=12.9Hz, 1H), 2.17 (qd, J=13.1, 12.3, 4.3Hz, 1H), 2.03-1.68 (m, 3H).

[0113] (S)-2-(1-acetylpiperidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] (S)-3-(4-nitro-1,3-dioxoisoindorin-2-yl)piperidine-1-ium chloride (0.16 g, 0.50 mmol) was added to a stirred suspension in DCM (12.5 mL) with triethylamine (0.17 mL, 2.4 equivalents) and acetyl chloride (0.08 mL, 2.3 equivalents). The solution was stirred for 1 hour. The reaction mixture was then concentrated on silica and purified by column chromatography. The compound was eluted using 1% MeOH in dichloromethane as the eluent, and the title compound was obtained as a colorless oil. Yield: 91% (0.15 g, 0.46 mmol). 1 H NMR (400MHz, CDCl3) δ 8.12(dd, J=8.5, 7.7Hz, 2H), 8.01-7.87(m, 1H), 4.73-4.61(m, 1H), 4.28-4.15(m, 1H), 3.90-3.72(m, 1H), 3.33(t, J=12.0H) z, 0.5H), 3.11(td, J=13.3, 2.6Hz, 0.5H), 2.61-2.23(m, 2H), 2.12(d, J=11.2Hz, 3H), 1.99-1.83(m, 2H), 1.70-1.50(m, 1H).

[0114] (S)-2-(1-acetylpiperidine-3-yl)-4-aminoisoindoline-1,3-dione(26): [ka] The compound was synthesized according to general procedure C under the following conditions: (S)-2-(1-acetylpiperidine-3-yl)-4-nitroisoindorin-1,3-dione (0.15 g, 0.46 mmol) and tin(II) chloride dihydrate (0.41 g, 4 equivalents) were stirred under reflux in ethyl acetate (30 mL) for 16 hours. The crude product was purified by column chromatography and eluted with phenylalanine as the eluent to obtain the title compound as a yellow solid. Yield: 93% (0.12 g, 0.43 mmol), obtained as a bright yellow solid. 1H NMR (400MHz, DMSO-D6) δ 7.42 (ddd, J=8.4, 7.0, 2.4Hz, 1H), 7.00-6.91 (m, 2H), 6.47 (br s, 2H), 4.44-4.33 (m, 1H), 3.98-3.75 (m, 2H), 3.58 (dd, J=12.9, 11.4Hz, 0.5H), 3.08 (t, J=11.9Hz, 0.5H), 2.97 (td, J=13.3, 2.6Hz, 0.5H), 2.43(td, J=13.1, 2.8Hz, 0.5H), 2.33-2.16(m, 1H), 2.00(d, J=22.8Hz, 3H), 1.89-1.68(m, 2H), 1.60-1.44(m, 0.5H), 1.43-1.29(m, 0.5H). 13 C NMR (101MHz, DMSO-D6) δ 169.29, 168.31, 168.22, 167.87, 146.51, 135.18, 132.16, 132.09, 121.40, 121.37, 110.68, 110. 64, 108.69, 47.98, 47.56, 46.68, 45.86, 43.11, 40.91, 27.59, 27.49, 25.27, 24.44, 21.35, 21.32. 1 H NMR (500MHz, DMSO-D6, 100℃) δ 7.43(dd, J=8.4, 7.0Hz, 1H), 7.01(d, J=8.4Hz, 1H), 6.97(d, J=7.0Hz, 1H), 6.24(s, 2H), 4 .00-3.87(m, 1H), 2.31(qd, J=12.6, 4.2Hz, 1H), 2.01(s, 3H), 1.89-1.78(m, 2H), 1.49(br s, 1H). LC-MS(ESI)m / z C 15 H 17 N3O3[M+H] + The calculated value is 287.1 and the measured value is 288.1. HPLC (method A): 7.99 points, purity 99%.

[0115] tert-ブチル(R)-3-(4-ニトロ-1,3-ジオキソイソインドリン-2-イル)ピペリジン-1-カルボキシレート:

change

[0116] (R)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)piperidine-1-ium chloride: [ka] To a stirred solution of tert-butyl(R)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)piperidine-1-carboxylate (0.77 g, 2.05 mmol) in 10 mL of siRNA, 4N HCl in 10 mL of dioxane was carefully added, and the mixture was stirred for 1 hour. The reaction mixture was then filtered, and the solid was washed with siRNA to obtain the title compound as a white solid. Yield: 75% (0.48 g, 1.54 mmol). 1H NMR (400MHz, DMSO-D6) δ 9.57 (br s, 1H), 9.40 (br s, 1H), 8.29(dd, J=8.0, 0.9Hz, 1H), 8.17(dd, J=7.5, 0.9Hz, 1H), 8.11-8.02(m, 1H), 4.47(tdd, J=12.3, 8.2, 4.0Hz, 1H) , 3.42-3.33(m, 3H), 3.29-3.21(m, 1H), 2.83(q, J=11.9Hz, 1H), 2.17(qd, J=13.1, 12.3, 4.3Hz, 1H), 1.96-1.73(m, 3H).

[0117] (R)-2-(1-acetylpiperidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] (R)-3-(4-nitro-1,3-dioxoisoindolin-2-yl)piperidine-1-ium chloride (0.16 g, 0.50 mmol) was added to a stirred suspension in DCM (12.5 mL) with triethylamine (0.17 mL, 2.4 equivalents) and acetyl chloride (0.08 mL, 2.3 equivalents). The solution was stirred for 1 hour. The reaction mixture was then diluted with dichloromethane and washed with 0.5 N HCl (aqueous solution) and brine. The organic fraction was dried over MgSO4, and the solvent was removed under reduced pressure to obtain the title compound as a colorless film. Yield: 99% (0.16 g, 0.50 mmol). 1 H NMR (400MHz, CDCl3) δ 8.12(dd, J=8.5, 7.7Hz, 2H), 8.03-7.89(m, 1H), 4.73-4.61(m, 1H), 4.30-4.13(m, 1H), 3.89-3.74(m, 1.5H), 3.33(t, J=12.0Hz, 0.5H), 3.10( td, J=13.3, 2.5Hz, 0.5H), 2.55(td, J=13.2, 3.0Hz, 0.5H), 2.50-2.31(m, 1H), 2.12(d, J=10.9Hz, 3H), 2.01-1.83(m, 2H), 1.71-1.50(m, 1H).

[0118] (R)-2-(1-acetylpiperidine-3-yl)-4-aminoisoindoline-1,3-dione(28): [ka] The compound was synthesized according to general procedure C under the following conditions: (R)-2-(1-acetylpiperidine-3-yl)-4-nitroisoindorin-1,3-dione (0.16 g, 0.50 mmol) and tin(II) chloride dihydrate (0.45 g, 4 equivalents) were stirred under reflux in ethyl acetate (30 mL) for 16 hours. The crude product was purified by column chromatography and eluted with ethyl acetate to obtain the title compound as a yellow solid. Yield: 95% (0.14 g, 0.47 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, DMSO-D6) δ 7.53-7.29(m, 1H), 7.05-6.83(m, 2H), 6.47(br s, 2H), 4.44-4.33(m, 1H), 3.99-3.77(m, 2H), 3.58(dd, J=12.9, 11.4Hz, 0.5H), 3.08(t, J=11.9Hz, 0.5H), 2.97(td, J=13.5, 2.6Hz, 0.5H), 2.43 (td, J=13.0, 2.8Hz, 0.5H), 2.26(qd, J=12.7, 4.2Hz, 1H), 2.00(d, J=22. 6Hz, 3H), 1.89-1.69(m, 2H), 1.63-1.43(m, 0.5H), 1.43-1.29(m, 0.5H). 13 C NMR (101MHz, DMSO-D6) δ 169.29, 168.31, 168.22, 167.87, 146.51, 135.18, 132.16, 132.09, 121.40, 121.37, 110.68, 110.64, 1 08.75, 108.69, 47.98, 47.56, 46.68, 45.86, 43.11, 40.91, 27.59, 27.49, 25.27, 24.44, 21.34, 21.31. 1H NMR (500MHz, DMSO-D6, 100℃) δ 7.43(dd, J=8.4, 7.0Hz, 1H), 7.01(d, J=8.4Hz, 1H), 6.97(d, J=7.0Hz, 1H), 6.24(br s, 2H), 4.01-3.88(m, 1H), 2.31(qd, J=12.3, 3.9Hz, 1H), 2.01(s, 3H), 1.90-1.78(m, 2H), 1.50(br s, 1H). LC-MS(ESI)m / z C 15 H 17 N3O3[M+H] + Calculated value: 287.1, measured value: 288.1. HPLC (Method A): 8.07 mins, purity 99%.

[0119] 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride: [ka] tert-butyl 3-(4-nitro-1,3-dioxoisoindolin-2-yl)azetidine-1-carboxylate (0.29 g, 0.83 mmol) was stirred at room temperature for 2 hours in a freshly prepared HCl solution of MeOH (2 M, 25 mL). After completion (TLC), the solvent was removed under reduced pressure, and the solid was washed with warmed MeOH. Yield: 67% (0.16 g, 0.55 mmol), obtained as pale yellow crystals. 1 H NMR (400MHz, DMSO-D6) δ 9.24(br s, 2H), 8.33 (d, J=8.0Hz, 1H), 8.22 (d, J=7.4Hz, 1H), 8.10 (t, J=7.8Hz, 1H), 5 .16-5.04(m, 1H), 4.51(dd, J=11.6, 7.3Hz, 2H), 4.22(dd, J=11.5, 9.1Hz, 2H).

[0120] 2-(1-acetylazetidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride (0.054 g, 0.19 mmol), triethylamine (0.05 mL, 2 equivalents), and acetic anhydride (0.02 mL, 1.3 equivalents) were stirred in dichloromethane (5 mL) at room temperature until complete. Yield: 92% (0.05 g, 0.18 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.20-8.13(m, 2H), 7.99(dd, J=8.3, 7.3Hz, 1H), 5.14(tt, J=8.7, 6.2Hz, 1H), 4.7 6(dd, J=8.6, 6.1Hz, 1H), 4.58-4.42(m, 2H), 4.36(t, J=9.2Hz, 1H), 1.95(s, 3H).

[0121] 2-(1-acetylazetidine-3-yl)-4-aminoisoindoline-1,3-dione(5): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-acetylazetidine-3-yl)-4-nitroisoindoline-1,3-dione (0.05 g, 0.17 mmol) and tin(II) chloride dihydrate (0.15 g, 4 equivalents) were stirred in ethyl acetate (20 mL) at 40°C for 16 hours. The product was purified by flash column chromatography using a C18 cartridge and eluted using a gradient of 0-50% CH3CN in H2O. Yield: 77% (0.03 g, 0.13 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, acetone-D6) δ 7.46(dd, J=8.4, 7.0Hz, 1H), 7.07-6.99(m, 2H), 5.04(tt, J=8.7, 6.1Hz, 1H), 4.7 1(dd, J=8.5, 6.0Hz, 1H), 4.52-4.41(m, 2H), 4.19(t, J=9.2Hz, 1H), 1.84(s, 3H). LC-MS(ESI)m / z C 13 H 13 N3O3[M+H] +Calculated value: 259.1, measured value: 260.0. HPLC (Method A): 6.54 mins, purity 99%.

[0122] 4-Nitro-2-(1-propionylazetidine-3-yl)isoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride (0.05 g, 0.18 mmol), triethylamine (0.05 mL, 2.2 equivalents), and propionyl chloride (0.02 mL, 1.2 equivalents) were stirred in dichloromethane (2 mL) at room temperature until completion. Yield: 86% (0.05 g, 0.15 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.20-8.12(m, 2H), 7.99(dd, J=8.3, 7.3Hz, 1H), 5.14(tt, J=8.6, 6.1Hz, 1H), 4.78-4.69(m , 1H), 4.56-4.41(m, 2H), 4.35(t, J=8.9Hz, 1H), 2.28-2.08(m, 2H), 1.16(t, J=7.5Hz, 3H).

[0123] 4-amino-2-(1-propionylazetidine-3-yl)isoindoline-1,3-dione(18): [ka] The compound was synthesized according to general procedure C under the following conditions: 4-nitro-2-(1-propionylazetidine-3-yl)isoindorin-1,3-dione (0.05 g, 0.15 mmol) and tin(II) chloride dihydrate (0.14 g, 4 equivalents) were stirred in ethyl acetate (5 mL) at 40°C for 72 hours. The product was purified by column chromatography and eluted with ethyl acetate. Yield: 98% (0.04 g, 0.15 mmol), obtained as a bright yellow solid. 1H NMR (400MHz, CDCl3) δ 7.42(dd, J=8.3, 7.1Hz, 1H), 7.13(dd, J=7.1, 0.7Hz, 1H), 6.89(dd, J=8.4, 0.7Hz, 1H), 5.04(tt, J=8. 7, 6.2Hz, 1H), 4.67-4.60(m, 4H), 4.36(t, J=9.0Hz, 2H), 2.19(q, J=7.6Hz, 2H), 1.17(t, J=7.5Hz, 3H). 13 C NMR (101MHz, CDCl3) δ 174.27, 169.67, 168.04, 145.87, 135.65, 132.22, 121.62, 112.91, 110.50, 38.30, 24.97, 9.01. LC-MS(ESI)m / z C 14 H 15 N3O3[M+H] + Calculated value: 273.1, measured value: 274.0. HPLC (Method A): 7.50 min, purity 99%.

[0124] 2-(1-butyrylazetidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride (0.05 g, 0.18 mmol), triethylamine (0.05 mL, 2.2 equivalents), and butyryl chloride (0.02 mL, 1.2 equivalents) were stirred in dichloromethane (2 mL) at room temperature until complete. Yield: 84% (0.05 g, 0.15 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.22-8.12(m, 2H), 7.99(dd, J=8.3, 7.3Hz, 1H), 5.14(tt, J=8.7, 6.2Hz, 1H), 4.74(dd, J=8.6, 6.2Hz, 1H), 4.55-4.41(m, 2H), 4.35(t, J=9.4Hz, 1H), 2.22-2.05(m, 2H), 1.70(p, J=7.1Hz, 2H), 0.98(t, J=7.4Hz, 3H).

[0125] 4-amino-2-(1-butyrylazetidine-3-yl)isoindoline-1,3-dione(23): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-butyrylazetidine-3-yl)-4-nitroisoindoline-1,3-dione (0.05 g, 0.15 mmol) and tin(II) chloride dihydrate (0.14 g, 4 equivalents) were stirred in ethyl acetate (5 mL) at 40°C for 72 hours. The product was purified by column chromatography and eluted with ethyl acetate. Yield: 97% (0.04 g, 0.15 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, CDCl3) δ 7.41(dd, J=8.3, 7.1Hz, 1H), 7.12(dd, J=7.1, 0.7Hz, 1H), 6.89(dd, J=8.4, 0.7Hz, 1H), 5.03(tt, J=8.7, 6.1Hz, 1H), 4.89(br s, 2H), 4.62(br s, 2H), 4.36(t, J=9.0Hz, 2H), 2.14(dd, J=8.4, 6.6Hz, 2H), 1.69(h, J=7.4Hz, 2H), 0.97(t, J=7.4Hz, 3H). 13 ¹³C NMR (101 MHz, methanol-D4) δ 175.67, 170.72, 169.63, 148.14, 136.37, 133.55, 122.55, 112.48, 110.73, 56.16, 53.93, 39.31, 34.27, 19.40, 14.06. LC-MS (ESI) m / z C 15 H 17 N3O3[M+H] + Calculated value: 287.1, measured value: 288.0. HPLC (Method A): 8.22 mins, purity 98%.

[0126] 2-(1-(3,3-dimethylbutanoyl)azetidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(4-nitro-1,3-dioxoisoindolin-2-yl)azetidine-1-ium chloride (0.10 g, 0.35 mmol), triethylamine (0.11 mL, 2.2 equivalents), and 3,3-dimethylbutyryl chloride (0.06 mL, 1.1 equivalents) were stirred in acetonitrile (2 mL) at room temperature until complete. Yield: 93% (0.11 g, 0.33 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.20-8.12(m, 2H), 7.99(dd, J=8.2, 7.3Hz, 1H), 5.11(tt, J=8.7, 6.2Hz, 1H), 4.79-4. 70(m, 1H), 4.56-4.41(m, 2H), 4.41-4.31(m, 1H), 2.05(d, J=5.0Hz, 2H), 1.08(s, 9H).

[0127] 4-Amino-2-(1-(3,3-dimethylbutanoyl)azetidine-3-yl)isoindoline-1,3-dione(24): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-(3,3-dimethylbutanoyl)azetidine-3-yl)-4-nitroisoindorin-1,3-dione (0.05 g, 0.15 mmol) and tin(II) chloride dihydrate (0.13 g, 4 equivalents) were stirred in ethyl acetate (10 mL) at 40°C for 72 hours. The product was purified by column chromatography and eluted using a gradient of 0-5% MeOH in dichloromethane. Yield: 98% (0.05 g, 0.14 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, CDCl3) δ 7.44-7.35(m, 1H), 7.10(dd, J=7.1, 0.7Hz, 1H), 6.88(dd, J=8.4, 0.7Hz, 1H), 5.00(tt , J=8.8, 6.2Hz, 1H), 4.87-4.54(m, 4H), 4.46-4.24(m, 2H), 2.04(s, 2H), 1.06(s, 9H). 13C NMR (101MHz, CDCl3) δ 172.36, 169.60, 168.07, 145.84, 135.62, 132.24, 121.60, 112.89, 110.53, 44.49, 37.83, 31.62, 29.97. LC-MS(ESI)m / z C 17 H 21 N3O3[M+H] + Calculated value: 315.2, measured value: 316.1. HPLC (Method A): 9.25 min, purity 96%.

[0128] 2-(1-(cyclopropanecarbonyl)azetidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride (0.07 g, 0.25 mmol), triethylamine (0.08 mL, 2.2 equivalents), and cyclopropanecarbonyl chloride (0.03 g, 1.2 equivalents) were stirred in acetonitrile (3 mL) at room temperature until completion. Yield: 78% (0.06 g, 0.19 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.20-8.13(m, 2H), 7.98(dd, J=8.2, 7.4Hz, 1H), 5.17(tt, J=8.7, 6.2Hz, 1H), 4.88(dd, J=8.5, 6.2Hz, 1H), 4.60(t, J=8.6Hz, 1H), 4.53(dd, J=10.0, 6.3Hz, 1H), 4.37(t, J=9.4Hz, 1H), 1.44(tt, J=7.9, 4.8Hz, 1H), 1.06-0.96(m, 2H), 0.86-0.74(m, 2H).

[0129] 4-Amino-2-(1-(cyclopropanecarbonyl)azetidine-3-yl)isoindoline-1,3-dione(19): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-(cyclopropanecarbonyl)azetidine-3-yl)-4-nitroisoindoline-1,3-dione (0.06 g, 0.19 mmol) and tin(II) chloride dihydrate (0.18 g, 4 equivalents) were stirred in ethyl acetate (10 mL) at 40°C for 48 hours. The product was purified by column chromatography and eluted with 1.5% MeOH in dichloromethane. Yield: 73% (0.04 g, 0.14 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, CDCl3) δ 7.42(dd, J=8.3, 7.1Hz, 1H), 7.13(dd, J=7.1, 0.7Hz, 1H), 6.88(dd, J=8.3, 0.7Hz, 1H), 5.08(tt, J=8.7, 6.2Hz, 1H), 4.76- 4.70(m, 4H), 4.44(t, J=9.0Hz, 2H), 1.44(tt, J=7.9, 4.6Hz, 1H), 1.01(dt, J=4.6, 3.3Hz, 2H), 0.79(dt, J=8.0, 3.4Hz, 2H). 13 C NMR (101MHz, CDCl3) δ 174.11, 169.75, 168.01, 145.80, 135.69, 132.37, 121.58, 113.02, 110.63, 53.92, 38.52, 10.45, 7.64. LC-MS(ESI)m / z C 15 H 15 N3O3[M+H] + Calculated value: 285.1, measured value: 286.0. HPLC (Method A): 7.70 min, purity 98%.

[0130] 2-(1-acryloylazetidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(4-nitro-1,3-dioxoisoindolin-2-yl)azetidine-1-ium chloride (0.07 g, 0.25 mmol), triethylamine (0.08 mL, 2.2 equivalents), and acryloyl chloride (0.02 mL, 1.2 equivalents) were stirred in acetonitrile (3 mL) until complete at room temperature. Yield: 46% (0.03 g, 0.11 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.20-8.12(m, 2H), 7.99(dd, J=8.3, 7.3Hz, 1H), 6.37(dd, J=17.0, 1.9Hz, 1H), 6.22(dd, J=17.0, 10.3Hz, 1H), 5.72 (dd, J=10.3, 1.9Hz, 1H), 5.18(tt, J=8.7, 6.2Hz, 1H), 4.87-4.79(m, 1H), 4.65-4.52(m, 2H), 4.44(t, J=9.1Hz, 1H).

[0131] 2-(1-acryloylazetidine-3-yl)-4-aminoisoindoline-1,3-dione(20): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-acryloylazetidine-3-yl)-4-nitroisoindorin-1,3-dione (0.03 g, 0.11 mmol) and tin(II) chloride dihydrate (0.10 g, 4 equivalents) were stirred in ethyl acetate (3 mL) at 40°C for 48 hours. The product was purified by column chromatography and eluted with 1.5% MeOH in dichloromethane. Yield: 82% (0.03 g, 0.09 mmol), obtained as a bright yellow solid. 1H NMR (400MHz, CDCl3) δ 7.41(dd, J=8.4, 7.1Hz, 1H), 7.12(d, J=7.0Hz, 1H), 6.92-6.85(m, 1H), 6.37(dd, J=17.0, 1.9Hz, 1H) , 6.22(dd, J=17.0, 10.3Hz, 1H), 5.70(dd, J=10.3, 1.9Hz, 1H), 5.08(tt, J=8.7, 6.2Hz, 1H), 4.71(br s, 2H), 4.53-4.4.45(m, 4H). 13 C NMR (101MHz, CDCl3) δ 169.57, 168.16, 165.75, 145.82, 135.72, 132.24, 127.95, 125.95, 121.63, 113.04, 38.53. LC-MS(ESI)m / z C 14 H 13 N3O3[M+H] + Calculated value: 271.1, measured value: 272.0. HPLC (Method A): 7.31 min, purity 99%.

[0132] 2-(1-benzoylazetidine-3-yl)-4-nitroisoindoline-1,3-dione: [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(4-nitro-1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride (0.05 g, 0.18 mmol), triethylamine (0.05 mL, 2.2 equivalents), and benzoyl chloride (0.03 mL, 1.2 equivalents) were stirred in dichloromethane (2 mL) at room temperature until complete. Yield: 99% (0.06 g, 0.17 mmol), obtained as a colorless solid. 1 H NMR (400MHz, CDCl3) δ 8.17-8.10(m, 2H), 7.96(dd, J=8.3, 7.3Hz, 1H), 7.72-7.63(m, 2H), 7.52-7.36(m, 3H), 5.18(tt, J=8.7, 6.1Hz, 1H), 4.91-4.83(m, 1H), 4.74-4.65(m, 1H), 4.62-4.53(m, 2H).

[0133] 4-amino-2-(1-benzoylazetidine-3-yl)isoindoline-1,3-dione(22): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-benzoylazetidine-3-yl)-4-nitroisoindorin-1,3-dione (0.06 g, 0.18 mmol) and tin(II) chloride dihydrate (0.16 g, 4 equivalents) were stirred in ethyl acetate (5 mL) at 40°C for 48 hours. The product was purified by column chromatography and eluted using a gradient of 50-75% ethyl acetate in pentane. Yield: 84% (0.05 g, 0.15 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, DMSO-D6) δ 7.69-7.62(m, 2H), 7.58-7.39(m, 4H), 6.97(dd, J=7.7, 6.2Hz, 2H), 6.51(br s, 2H), 5.00(tt, J=8.6, 5.8Hz, 1H), 4.72-4.57(m, 2H), 4.50(dd, J=10.2, 5.9Hz, 1H), 4.34(t, J=9.5Hz, 1H). LC-MS(ESI)m / z C 18 H 15 N3O3[M+H] + Calculated value: 321.1, measured value: 322.0. HPLC (Method A): 8.91 min, purity 99%.

[0134] tert-butyl(1-acetylazetidine-3-yl)carbamate: [ka] 3-N-Boc-amino-azetidine hydrochloride (2.0 g, 9.58 mmol) was suspended in dichloromethane (40 mL), and the suspension was cooled to 0°C using an ice bath. Triethylamine (4.0 mL, 3 equivalents) was added, followed by dropwise addition of acetyl chloride (1.02 mL, 1.5 equivalents in 10 mL of dichloromethane). After the addition was complete (TLC), the reaction mixture was warmed to room temperature and maintained at that temperature for 1.5 hours. H2O was added, the layers were separated, the organic fraction was washed with brine, and dried over MgSO4. The crude product was purified by column chromatography, first eluted with 50 / 50 ethyl acetate in pentane, and then with 100% ethyl acetate. Yield: 83% (1.71 g, 8.0 mmol), obtained as a colorless oil. 1 H NMR (400MHz, CDCl3) δ 6.07(br s, 1H), 4.43-4.18(m, 2H), 4.16-4.03(m, 1H), 3.97-3.82(m, 1H), 3.78-3.65(m, 1H), 1.71(s, 3H), 1.30(s, 9H).

[0135] 1-Acetylazetidine-3-aminium chloride: [ka] 1.71 g, 8.0 mmol of tert-butyl(1-acetylazetidine-3-yl)carbamate was placed in 40 mL of DCM, and 27 mL of HCl (4.0 M in dioxane) was carefully added. The solution was stirred at room temperature for 1 hour, and then 40 mL of diethyl ether was added to obtain a precipitate of the product. The flask was left overnight to complete the precipitation, and then the solid was filtered and washed with ether. Yield: 99% (1.21 g, 8.0 mmol), obtained as a grayish-white solid. 1 H NMR (400MHz, DMSO-D6) δ 8.81(s, 3H), 4.37-4.28(m, 1H), 4.12(dd, J=9.6, 4.5Hz, 1H), 4.06-3.88(m, 2H), 3.84(dd, J=9.9, 4.4Hz, 1H), 1.75(s, 3H).

[0136] 2-(1-acetylazetidine-3-yl)-5-nitroisoindoline-1,3-dione: [ka] Step 1: Triethylamine (0.11 mL, 1.2 equivalents) was added to a stirred suspension of 5-nitroisobenzofuran-1,3-dione (0.13 g, 0.66 mmol) and 1-acetylazetidine-3-aminium chloride (0.15 g, 1.5 equivalents) in THF (5 mL). Immediate precipitation was induced upon addition of triethylamine. The suspension was stirred at room temperature for 4 hours. The reaction mixture was filtered, the solid was washed with THF, and then dried under reduced pressure. Step 2: The solid was suspended in acetic anhydride (5 mL), and then sodium acetate trihydrate (0.08 g, 1 equivalent) was added. The reaction mixture was heated to 80°C and maintained for 40 minutes, after which complete conversion was shown by TLC. The reaction mixture was poured onto ice and extracted with ethyl acetate (3 x 50 mL). The combined organic fraction was dried over MgSO4, and then the solvent was removed under reduced pressure. Yield: 41% (0.07 g, 0.25 mmol) was obtained as a light brown solid through two processes. 1 H NMR (400MHz, CDCl3) δ 8.69-8.59(m, 2H), 8.07(dd, J=8.1, 0.7Hz, 1H), 5.11(tt, J=8.7, 6.1Hz, 1H), 4.72(t, J=7.6Hz, 1H), 4.60-4.39(m, 2H), 4.33(t, J=9.6Hz, 1H), 1.92(s, 3H).

[0137] 2-(1-acetylazetidine-3-yl)-5-aminoisoindoline-1,3-dione(6): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-acetylazetidine-3-yl)-5-nitroisoindoline-1,3-dione (0.07 g, 0.25 mmol) and tin(II) chloride dihydrate (0.22 g, 4 equivalents) were stirred in ethyl acetate (12 mL) at 40°C for 24 hours. The product was purified by column chromatography and eluted with 2% MeOH in chloroform. Yield: 62% (0.04 g, 0.16 mmol), obtained as a bright yellow solid. 1 H NMR (400MHz, DMSO-D6) δ 7.50(d, J=8.2Hz, 1H), 6.92(d, J=2.0Hz, 1H), 6.80(dd, J=8.2, 2.1Hz, 1H), 6.52(br s, 2H), 4.90(tt, J=8.7, 6.0Hz, 1H), 4.52(dd, J=8.6, 6.0Hz, 1H), 4.37(td, J=8. 7, 0.9Hz, 1H), 4.26(dd, J=9.7, 6.1Hz, 1H), 4.09(t, J=9.2Hz, 1H), 1.79(s, 3H). 13 C NMR (101MHz, DMSO-D6) δ 169.47, 168.03, 167.55, 155.13, 134.34, 125.04, 116.81, 116.37, 106.92, 54.71, 52.39, 37.66, 18.87. LC-MS(ESI)m / z C 13 H 13 N3O3[M+H] + Calculated value: 259.1, measured value: 260.1. HPLC (Method B): 9.13 mins, purity 99%.

[0138] 5-(1-acetylazetidine-3-yl)-4H-thieno[2,3-c]pyrrole-4,6(5H)-dione(9): [ka] Step 1: Triethylamine (0.18 mL, 2 equivalents) was added to a stirred suspension of thieno[2,3-c]furan-4,6-dione (0.09 g, 0.9 equivalents) and 1-acetylazetidine-3-aminium chloride (0.1 g, 0.66 mmol) in THF (5 mL). Immediate precipitation was induced upon addition of triethylamine. The suspension was stirred overnight at room temperature, and then the mixture was concentrated under reduced pressure. Step 2: The crude product was suspended in acetic anhydride (5 mL), and then sodium acetate trihydrate (0.10 g, 1 equivalent) was added. The reaction mixture was heated to 80°C and maintained for 1 hour, after which complete conversion was shown by LC-MS. The reaction mixture was concentrated under reduced pressure, and the remaining acetic anhydride was removed by co-evaporation with toluene (3x). The crude product was purified by flash column chromatography and eluted using a gradient of 0-5% MeOH in chloroform. Yield: 42% (0.07 g, 0.28 mmol) was obtained as a white solid through two steps. 1 H NMR (400MHz, CDCl3) δ 7.86(d, J=4.7Hz, 1H), 7.35(d, J=4.7Hz, 1H), 5.03(tt, J=8.7, 6.3Hz, 1H), 4.73(dd, J=8.6, 6.1 Hz, 1H), 4.51(dd, J=10.0, 6.4Hz, 1H), 4.43(t, J=8.6, 1H), 4.33(t, J=9.4Hz, 1H), 1.96(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.60, 163.16, 162.03, 144.38, 140.61, 138.50, 121.39, 55.02, 53.04, 38.74, 19.04. LC-MS(ESI)m / z C 11 H 10 N2O3S [M+H] + Calculated value: 250.0, measured value: 251.0. HPLC (Method B): 9.31 min, purity 99%.

[0139] N-(2-(1-acetylazetidine-3-yl)-1,3-dioxoisoindoline-4-yl)acetamide(7): [ka] 2-(1-acetylazetidine-3-yl)-4-nitroisoindoline-1,3-dione (0.1 g, 0.33 mmol) was stirred in 10 mL of toluene (10 mL), to which tin(II) chloride dihydrate (0.3 g, 4 equivalents) was added. The solution was then maintained under reflux for 1.5 hours, after which complete conversion to aniline was shown by LC / MS. The reaction mixture was cooled to room temperature, and acetic anhydride (0.3 mL, 10 equivalents) was carefully added. The mixture was then heated and maintained under reflux for a further 1.5 hours, after which the reaction mixture was cooled to room temperature, diluted with toluene, and quenched by adding a saturated aqueous solution of NaHCO3. The layers were separated, and the aqueous fraction was extracted once from toluene. The combined organic fraction was washed with brine and dried over MgSO4. The crude product was purified by column chromatography, and the title compound was obtained as a pale yellow solid by elution using a gradient of 0-3% MeOH in dichloromethane. Yield: 64% (0.07g, 0.22 mmol). 1 H NMR (400MHz, CDCl3) δ 9.43(s, 1H), 8.75(d, J=8.3Hz, 1H), 7.66(dd, J=8.4, 7.5Hz, 1H), 7.48(dd, J=7.3, 0.7Hz, 1H), 5.03(tt, J=8.7, 6.2Hz, 1H) , 4.72-4.64(m, 1H), 4.48(dd, J=9.7, 6.4Hz, 1H), 4.41(t, J=8.6Hz, 1H), 4.31(t, J=9.4Hz, 1H), 2.25(s, 3H), 1.92(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.61, 169.69, 169.33, 167.02, 137.75, 136.43, 130.97, 125.17, 118.30, 115.08, 54.77, 52.78, 38.36, 25.03, 19.06. LC-MS(ESI)m / z C 15 H 15 N3O4[M+H] + Calculated value: 301.1, measured value: 302.1. HPLC (Method B): 9.97 mins, purity 95.1%.

[0140] 5-Chloro-2-methyl-3-nitrobenzoic acid: [ka] At -10°C, 5-chloro-2-methylbenzoic acid (5.0 g, 29.3 mmol) was dissolved in sulfuric acid (34 mL) in a divided manner. After dissolving the substance, a 2:1 mixture of sulfuric acid and nitric acid (65%) (8.5 mL) was added dropwise while maintaining the temperature between -10°C and 0°C. After the addition, the reaction mixture was stirred for several minutes, then poured onto ice and subsequently extracted with ethyl acetate. The organic fraction was dried over MgSO4, the solvent was evaporated under reduced pressure, and the crude product was recrystallized from warmed methanol to obtain the title compound as a pale yellow solid. Yield: 37% (2.30 g, 10.7 mmol). 1 H NMR (400MHz, CDCl3) δ 11.44(s, 1H), 8.19(d, J=2.3Hz, 1H), 7.90(d, J=2.4Hz, 1H), 2.67(s, 3H).

[0141] 5-chloro-3-nitrophthalic acid: [ka] 5-Chloro-2-methyl-3-nitrobenzoic acid (2.16 g, 10 mmol) was suspended in H2O (40 mL), and then sodium hydroxide (0.8 g, 2 equivalents) was added. The solution was then stirred at 85°C, and potassium permanganate (3.16 g, 2 equivalents) was added in installments over 3 hours. After the addition was complete, the reaction mixture was maintained at 85°C for 1 hour. The reaction mixture was cooled to room temperature, filtered, and the solid was washed with warmed H2O (2 x 30 mL). The filtrate was acidified to pH=1-2 with 2N HCl (aqueous solution), pharmaceutically acceptable, and the crude product was extracted (3 x 100 mL). The combined organic fraction was dried over MgSO4 and concentrated under reduced pressure. The product was then purified by column chromatography, first eluted with 99:1:1 DCM / MeOH / AcOH, and then with 48:48:4 DCM / MeOH / AcOH to obtain the title phthalic acid as a yellow solid. Yield: 46% (1.13 g, 4.60 mmol).

[0142] 2-(1-acetylazetidine-3-yl)-6-chloro-4-nitroisoindoline-1,3-dione: [ka] Step 1: 5-chloro-3-nitrophthalic acid (0.15 g, 0.61 mmol) and sodium acetate trihydrate (0.08 g, 1 equivalent) were added to acetic anhydride (10 mL) and stirred at 80°C for 1 hour. The solvent was then removed under reduced pressure, and the remaining acetic anhydride was removed by co-evaporation with toluene (3x). The crude phthalic anhydride was suspended in THF (4 mL), and then 1-acetylazetidine-3-aminium chloride (0.1 g, 1.1 equivalents) and triethylamine (0.17 mL, 2 equivalents) were added. The resulting suspension was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure and further processed without any purification. Step 2: Crude 2-((1-acetylazetidine-3-yl)carbamoyl)-4-chloro-6-nitrobenzoic acid was added to acetic anhydride (5 mL), and sodium acetate trihydrate (0.08 g, 1 equivalent) was added. This solution was heated to 80°C and maintained at that temperature for 1 hour. LC / MS showed complete conversion from benzoic acid to phthalimide. The reaction mixture was concentrated under reduced pressure, co-evaporated with toluene (2x), and finally purified by flash column chromatography. The title compound was eluted using a gradient of 0-10% MeOH in DCM to obtain a pale yellow oily substance. Yield: 27% (0.05 g, 0.15 mmol) across both steps. 1 H NMR (400MHz, CDCl3) δ 8.14(d, J=1.7Hz, 1H), 8.12(d, J=1.7Hz, 1H), 5.13(tt, J=8.7, 6.1Hz, 1H), 4.74(dd , J=8.8, 6.2Hz, 1H), 4.55-4.41(m, 2H), 4.36(dd, J=10.2, 8.7Hz, 1H), 1.95(s, 3H).

[0143] 2-(1-acetylazetidine-3-yl)-4-amino-6-chloroisoindoline-1,3-dione(10): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-acetylazetidine-3-yl)-6-chloro-4-nitroisoindoline-1,3-dione (54 mg, 0.17 mmol) was dissolved in ethyl acetate (10 mL), and then tin(II) chloride dihydrate (0.35 g, 9 equivalents) was added. The resulting solution was then stirred under reflux for 8 hours. The title compound was purified by flash chromatography and eluted using a gradient of 0-5% MeOH in chloroform to obtain a bright yellow solid. Yield: 92% (0.05 g, 0.15 mmol). 1 H NMR (400MHz, CDCl3) δ 7.07(s, 1H), 6.90(s, 1H), 5.50(br s, 2H), 5.07-4.95(m, 1H), 4.69(t, J=7.4Hz, 1H), 4.57-4.45(m, 1H), 4.40(t, J=8.6Hz, 1H), 4.30(t, J=9.4Hz, 1H), 1.93(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.80, 168.94, 166.94, 146.39, 141.98, 133.71, 120.54, 113.50, 109.20, 54.94, 52.91, 38.24, 19.14. LC-MS(ESI)m / z C 13 H 12 ClN3O3[M+H] + Calculated value: 293.1, measured value: 294.0. HPLC (Method A): 7.88 mins, purity 95%.

[0144] 3-chloro-6-nitrophthalic acid: [ka] At 0°C, 2.5 g (13.7 mmol) of 3-chlorophthalic anhydride was added in fractional amounts to a stirred solution of nitric acid and concentrated sulfuric acid (1:2, 9 mL). After addition, the suspension was warmed to room temperature and maintained at that temperature for 3 hours. The reaction mixture was then cooled again to 0°C, and ice was added to the mixture. The slurry was filtered, and the filtrate was extracted with ethyl acetate (3x). The combined organic fraction was dried over MgSO4, the solvent was removed under reduced pressure, and the crude product was further purified by column chromatography. Elution was performed using 5% MeOH in dichloromethane to obtain the title phthalic acid as a grayish-white solid. Yield: 56% (1.87 g, 7.61 mmol). 1 H NMR (400MHz, DMSO-D6) δ 14.39 (br s, 2H), 8.16 (d, J=8.7Hz, 1H), 7.93 (d, J=8.7Hz, 1H).

[0145] 2-(1-acetylazetidine-3-yl)-4-chloro-7-nitroisoindoline-1,3-dione: [ka] Step 1: 3-chloro-6-nitrophthalic acid (0.2 g, 0.81 mmol) and sodium acetate trihydrate (0.11 g, 1 equivalent) were added to acetic anhydride (5 mL) and stirred at 80°C for 1 hour. The resulting suspension was concentrated under reduced pressure, and the remaining acetic anhydride was removed by co-evaporation with toluene (3x). Next, the crude phthalic anhydride was added to THF / H2O (1:1, 25 mL), to which 1-acetylazetidine-3-aminium chloride (0.13 g, 1.06 equivalents) and triethylamine (0.23 mL, 2 equivalents) were added. The solution was stirred at room temperature for 2 hours, and then the solvent was removed under reduced pressure, and the remaining water was removed by co-evaporation with toluene. Step 2: Crude 2-((1-acetylazetidine-3-yl)carbamoyl)-3-chloro-6-nitrobenzoic acid was suspended in acetic anhydride (5 mL), and sodium acetate trihydrate (0.11 g, 1 equivalent) was added. The resulting solution was stirred at 80°C for approximately 45 minutes. LC / MS showed complete conversion to the desired phthalimide, so the reaction mixture was cooled to room temperature, then diluted with water, and the remaining acetic anhydride was carefully quenched by titration with saturated aqueous solution of NaHCO3. Ethyl acetate was then added, and the product was extracted (3x). The combined organic fraction was dried over MgSO4 and concentrated under reduced pressure. Finally, the title compound was purified by column chromatography and eluted with 2% MeOH in DCM to obtain a white solid. Yield: 19% (0.05 g, 0.15 mmol). 1 H NMR (400MHz, CDCl3) δ 8.05(d, J=8.6Hz, 1H), 7.87(d, J=8.7Hz, 1H), 5.12(qt, J=8.9, 6.2Hz, 1H), 4.79-4.71(m, 1H), 4.57-4.30(m, 3H), 1.95(d, J=1.6Hz, 3H).

[0146] 2-(1-acetylazetidine-3-yl)-4-amino-7-chloroisoindoline-1,3-dione(11): [ka] The compound was synthesized according to general procedure C under the following conditions: 2-(1-acetylazetidine-3-yl)-4-chloro-7-nitroisoindoline-1,3-dione (0.05 g, 0.15 mmol) and tin(II) chloride dihydrate (0.20 g, 6 equivalents) were added to ethyl acetate (10 mL), stirred at reflux temperature for 6 hours, and then LC / MS and TLC demonstrated complete conversion to the desired aniline. The crude product was purified by column chromatography and eluted with 1.5% MeOH in chloroform to obtain the title compound as a bright yellow solid. Yield: 75% (0.03 g, 0.11 mmol). 1 H NMR (400MHz, CDCl3) δ 7.34(d, J=8.8Hz, 1H), 6.86(d, J=8.8Hz, 1H), 5.44(s, 2H), 5.06(tt, J=8.7, 6.2Hz, 1H), 4.74(dd, J=8.4, 6 .2Hz, 1H), 4.53(dd, J=10.0, 6.3Hz, 1H), 4.41(td, J=8.6, 1.0Hz, 1H), 4.32(t, J=9.4Hz, 1H), 1.94(s, 3H). 13 C NMR (101MHz, CDCl3) δ 214.52, 170.71, 165.64, 144.71, 137.44, 127.10, 123.08, 119.36, 54.81, 52.89, 38.22, 19.15. LC-MS(ESI)m / z C 13 H 12 Calculated value of ClN3O3 [M+H] + 293.1, measured value 294.0. HPLC (Method A): 7.53 mins, purity 98.6%.

[0147] 4-methyl-3-nitrophthalic acid: [ka] 4-methylphthalic acid (2.51 g, 13.9 mmol) was added in fractions to a stirred solution of nitric acid and concentrated sulfuric acid (1:2, 9 mL) at 0°C. The suspension was stirred at 0°C for 15 minutes, and then allowed to reach room temperature over 2 hours. The reaction mixture was then poured onto ice, the solid was filtered off, and washed with cold water. The combined filtrate was extracted with ethyl acetate (2x), and the combined organic fraction was dried over MgSO4. The solvent was removed under reduced pressure, and the resulting solid was combined with the precipitated solid to obtain a 60 / 40 mixture of 4-methyl-5-nitrophthalic acid and 4-methyl-3-nitrophthalic acid as white solids. Yield: 70% (2.20 g, 10.0 mmol).

[0148] 2-(1-acetylazetidine-3-yl)-5-methyl-4-nitroisoindoline-1,3-dione: [ka] Step 1: A mixture of 4-methyl-5-nitrophthalic acid and 4-methyl-3-nitrophthalic acid (2.2 g, 10.0 mmol) was added to acetic anhydride (10 mL) and stirred at 90°C for 4 hours. The resulting clear solution was then cooled to room temperature and concentrated under reduced pressure. The remaining acetic anhydride was removed by co-evaporation with toluene (3x). Next, the intermediate anhydride (0.3 g, 1.45 mmol) was added to THF (5 mL), to which 1-acetylazetidine-3-aminium chloride (0.22 g, 1 equivalent), triethylamine (1 mL, 5 equivalents), and H2O (5 mL) were added. The resulting solution was stirred at room temperature for 1 hour, then the THF was removed under reduced pressure, and the reaction mixture was acidified to pH=1 by titration with HCl (aqueous solution, 2.0 M). The product was extracted with ELISA (2x), and the combined organic fraction was dried over MgSO4, and the solvent was evaporated to dryness under reduced pressure. Step 2: The intermediate was placed in acetic anhydride (10 mL) and stirred at 90°C for 1 hour. Complete conversion to the desired phthalimide was then demonstrated by LC / MS. The reaction mixture was concentrated on silica and purified by column chromatography, and eluted using a gradient of 1-2% MeOH in DCM. The title compound was isolated as a colorless oil as a 1:1 mixture of 2-(1-acetylazetidine-3-yl)-5-methyl-4-nitroisoindoline-1,3-dione and 2-(1-acetylazetidine-3-yl)-5-methyl-6-nitroisoindoline-1,3-dione. Yield: 25% (0.11 g, 0.36 mmol). 1 H NMR (400MHz, CDCl3) δ 8.33(s, 1H), 7.92(d, J=7.7Hz, 1H), 7.89(s, 1H), 7.76(dd, J=7.7, 0.5Hz, 1H), 5.31(s, 1H), 5.11(dtt, J=15.0, 8.7, 6.2Hz , 2H), 4.75(dt, J=8.7, 6.0Hz, 2H), 4.63-4.30(m, 6H), 3.95-3.87(m, 1H), 2.74(s, 3H), 2.49(s, 3H), 1.97(d, J=8.5Hz, 6H).

[0149] 2-(1-acetylazetidine-3-yl)-4-amino-5-methylisoindoline-1,3-dione(12): [ka] The compound was synthesized according to general procedure C under the following conditions: A mixture of nitro-substituted methyl phthalimide (0.11 g, 0.36 mmol, isomer ratio approximately 1:1) and tin(II) chloride dihydrate (0.33 g, 4 equivalents) was placed in ethyl acetate (15 mL) and stirred at reflux temperature for 4 hours. The crude product was purified by column chromatography, and the title compound was obtained as a bright yellow solid by elution using a 1-2% MeOH gradient in chloroform. Yield: 45% (0.02 g, 0.08 mmol). 1 H NMR (400MHz, CDCl3) δ 7.31(d, J=7.4Hz, 1H), 7.08(d, J=7.3Hz, 1H), 5.46-4.82(m, 2H), 5.03(tt, J=8.7, 6.2Hz, 1H), 4.81-4.66(m, 1H), 4.62-4.47(m, 1H), 4.46-4.28(m, 2H), 2.24(s, 3H), 1.94(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.74, 170.03, 168.03, 144.45, 135.89, 130.05, 113.01, 110.49, 55.02, 52.96, 38.07, 19.12, 17.18. LC-MS(ESI)m / z C 14 H 15 Calculated value of N3O3[M+H]+: 273.1, measured value: 274.0. HPLC (Method A): 7.31 mins, purity 96.9%.

[0150] 2-(1-acetylazetidine-3-yl)-5-amino-6-methylisoindoline-1,3-dione(13): [ka] The compound was synthesized according to general procedure C under the following conditions: A mixture of nitro-substituted methyl phthalimide (0.11 g, 0.36 mmol, isomer ratio approximately 1:1) and tin(II) chloride dihydrate (0.33 g, 4 equivalents) was placed in ethyl acetate (15 mL) and stirred at reflux temperature for 4 hours. The crude product was purified by column chromatography, and the title compound was obtained as an orange-yellow solid by elution using a 1-2% MeOH gradient in chloroform. Yield: 55% (0.03 g, 0.10 mmol). 1 H NMR (400MHz, CDCl3) δ 7.49(s, 1H), 7.05(s, 1H), 5.02(tt, J=8.7, 6.2Hz, 1H), 4.76-4.68(t, J=7.2Hz, 1H), 4.62-4.23(m, 5H), 2.24(s, 3H), 1.93(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.67, 168.44, 168.18, 150.97, 132.35, 126.91, 126.05, 120.16, 108.33, 55.03, 53.05, 38.16, 19.16, 18.08. LC-MS(ESI)m / z C 14 H 15 Calculated value of N3O3[M+H]+: 273.1, measured value: 274.0. HPLC (Method A): 6.87 mins, purity 97.7%.

[0151] Ethyl 1,3-dioxoisoindoline-2-carboxylate: [ka] At 0°C, ethyl chloroformate (5.76 mL, 1.2 equivalents) was added dropwise to a stirred solution of phthalimide (7.36 g, 50 mmol) and triethylamine (9.0 mL, 1.3 equivalents) in DMF (25 mL). After the addition was complete (20 minutes), the resulting suspension was stirred at room temperature for 1.5 hours. The reaction mixture was then poured onto ice, the resulting white precipitate was filtered off, washed with water, and then dried to obtain the title compound as a white solid. Yield: 87% (9.0 g, 43.6 mmol). 1H NMR (400MHz, CDCl3) δ 7.98 (dd, J=5.5, 3.1Hz, 2H), 7.84 (dd, J=5.6, 3.0Hz, 2H), 4.50 (q, J=7.1Hz, 2H), 1.45 (t, J=7.1Hz, 3H).

[0152] tert-butyl 3-(1,3-dioxoisoindorin-2-yl)azetidine-1-carboxylate: [ka] The compound was synthesized according to general procedure B under the following conditions: tert-butyl 3-aminoazetidine-1-carboxylate (1.0 g, 581 mmol), ethyl 1,3-dioxoisoindoline-2-carboxylate (1.27 g, 1 equivalent), and triethylamine (1.62 ml, 2 equivalents) were placed in THF (20 mL), and the resulting solution was stirred for 16 hours. The solvent was then removed under reduced pressure, and the crude product was placed in SiO2 and washed with HCl (0.5 M, aqueous solution) and brine. The organic fraction was dried over MgSO4 and concentrated under reduced pressure. The title compound was then obtained as a white solid by column purification using elution with 0-1% MeOH in DCM. Yield: 95% (1.67 g, 5.52 mmol). 1 H NMR (400MHz, CDCl3) δ 7.91-7.82(m, 2H), 7.80-7.70(m, 2H), 5.09-4.97(m, 1H), 4.53(dd, J=8.9, 6.3Hz, 2H), 4.24(t, J=8.9Hz, 2H), 1.49(s, 9H).

[0153] 3-(1,3-dioxoisoindoline-2-yl)azetidine-1-ium chloride: [ka] tert-butyl 3-(1,3-dioxoisoindolin-2-yl)azetidine-1-carboxylate (1.76 g, 5.82 mmol) was stirred in a solution of HCl in MeOH (2.0 M, 30 mL). After 2 hours, complete conversion was observed by TLC. The solvent was removed under reduced pressure, and the remaining HCl was removed by co-evaporation with toluene (3x) to obtain the title compound as a grayish-white solid. Yield: 99% (1.40 g, 5.82 mmol). 1 H NMR (400MHz, DMSO-D6) δ 10.06-8.81(m, 2H), 7.95-7.83(m, 4H), 5.15-5.02(m, 1H), 4.60-4.51(m, 2H), 4.22-4.08(m, 2H).

[0154] 2-(1-acetylazetidine-3-yl)isoindoline-1,3-dione(3): [ka] The compound was synthesized according to general procedure E under the following conditions: 3-(1,3-dioxoisoindorin-2-yl)azetidine-1-ium chloride (0.35 g, 1.45 mmol), acetyl chloride (0.12 mL, 1.2 equivalents), and triethylamine (0.42 mL, 2.1 equivalents) were stirred in DCM (10 mL) for 1 hour. The title compound was obtained as a white solid by column purification using 0-2% MeOH in DCM and subsequent recrystallization from a hexane mixture in toluene. Yield: 22% (0.08 g, 0.32 mmol). 1 H NMR (400MHz, CDCl3) δ 7.90-7.81(m, 2H), 7.80-7.70(m, 2H), 5.09(tt, J=8.7, 6.2Hz, 1H), 4.81-4.62(m, 1H), 4.62-4.47(m, 1H), 4.47-4.24(m, 2H), 1.94(s, 3H). 13 C NMR (101MHz, CDCl3) δ 170.66, 167.80, 153.51, 134.61, 131.66, 123.71, 119.93, 38.45, 19.10. LC-MS(ESI)m / z C 13 H 12Calculated value of N2O3[M+H]+: 244.3, measured value: 245.1. HPLC (Method B): 9.77 mins, purity 99%.

[0155] material and method Mouse model Cdh5(PAC)-CRE ERT2 ;Eng f / f The mice were provided by Professor HM. Arthur (Biosciences Institute, Faculty of Medical Sciences, Newcastle University, UK). All experiments were maintained on the C57BL6 / J genetic background.

[0156] The C57BL6 / JRj mice were provided by Janvier Lab.

[0157] Eng-iKO e For this study, 8-12 week old male and female mice were administered tamoxifen (2.5 mg per day, total volume 50 ml per infusion) by intraperitoneal infusion for 5 consecutive days to deplete endoglin in endothelial cells. Tamoxifen was purchased from Sigma-Aldrich (catalog number T5648) and dissolved in a 1:4 ethanol (Merck, catalog number 107017) / coil oil (Sigma-Aldrich, catalog number C8267) vehicle. Intraperitoneal infusion of either the vehicle or thalidomide or its analogue was started for 2 consecutive days starting on day 6 after endoglin depletion (day 0 corresponds to the day of the first tamoxifen infusion), and the animals were collected on day 9.

[0158] For irradiation of the head, including the eyes, the animals were fixed in a containment device without anesthesia. The heads of adult mice were exposed to a Philips / YXYLON MG325 X-ray generator (voltage 200 kV, current 21 mA). The X-ray radiation was sighted by a lead block placed 50 cm from the source. Four mice were irradiated per session. The total dose of 20 Gy was irradiated as a single fraction at a dose rate of 0.9 Gy / min. The mice were given a vehicle (DMSO), thalidomide, or lenalidomide (concentration: 200 mmol / kg) -1 Either a total volume of 50 ml per infusion or one of the following was intraperitoneally injected on day -2, day -1, day +1, day +2, day +5, day +6, and day +7, relative to day 0, when single-fractionated X-ray ionizing radiation was received. Mice were harvested on day +8, day +90, or day +180 after receiving a single dose of radiotherapy on day 0.

[0159] Perisite patch clamp records The patch clamp experiment was carried out as described. 4 In short, fresh mouse retinas were dissected and perfused with bicarbonate buffer containing 124 mM NaCl, 26 mM NaHCO3, 1 mM NaH2PO4, 2.5 mM KCl, 1 mM MgCl2, 2.5 mM CaCl2, and 10 mM glucose, then bubbling with 95% O2 / 5% CO2 and maintaining pH 7.3 at room temperature. Electrodes were filled with the same bicarbonate buffer and used to stimulate pericytes by pressing the electrodes against the cell bodies and applying voltage pulses (0.02 ms pulses, 10 Hz for 5 seconds), providing current intensities ranging from 1 to 3.5 mA. Pipette resistance was measured before each experiment to specify the input voltage. Pericytes were stimulated by applying voltage pulses 20 seconds after patch clamping, and vasoconstriction was recorded for 210 seconds, acquiring a total of 300 images.

[0160] Vascular permeability experiment The vascular permeability experiment was conducted as described. 5 In short, lysine-immobilizable cadaverine (5 mg / ml in saline) conjugated to Alexa Fluor-555.-1 Cadaverine Alexa Fluor-555 (Invitrogen, catalog number A30677) was intravenously injected into the tail vein of 12-week-old adult C57BL6 / JRj mice. The circulation time of cadaverine Alexa Fluor-555 was 2 hours. Animals were recovered by perfusion with HEPES buffered saline for fluorescence tracer quantification or by perfusion with 4% PFA for whole-brain imaging. For fluorescence quantification, the amount of injected cadaverine Alexa Fluor-555 was 120 mg per 20 grams of body weight. Brains were extracted and homogenized in 1% Triton X-100 in PBS pH 7.2. Brains were centrifuged at 16,000 rpm for 20 minutes, and the relative fluorescence of the supernatant was measured using a POLARstar Omega (BMG Labtech) fluorometer (ex / em 544 / 590 nm).

[0161] For whole-brain imaging, the injected doses of Alexa Fluor-555 or BSA Alexa Fluor-555 were 500 mg or 1850 mg per 20 grams of body weight, respectively. The brains were excised, fixed overnight in 4% PFA, and images were acquired using an Axio Zoom.V16 fluorescence stereomicroscope (Zeiss). Images were acquired with exposure times that did not saturate the fluorescence signals originating from the brains of irradiated (RT) animals.

[0162] Antibody and immunofluorescence staining All images were acquired using a Leica SP5 confocal microscope (Leica Microsystems). Brightness and contrast were adjusted using ImageJ, and the images are presented as maximum intensity projections.

[0163] For each image, the endothelial cell surface was defined as a Glut1(+) pixel and determined by the total gray value.

[0164] To quantify the number of neurons, we reconstructed the 25 micron maximum projection z stack and previously described 6As described above, the number of NeuN-positive neurons per mm² was determined using the Cell Counter plugin analysis tool in ImageJ software. In each animal, six fields (388.74 x 388.74 mm) randomly selected from the primary somatosensory barrel cortex were analyzed in three non-adjacent sections (approximately 250 mm apart).

[0165] statistical analysis Statistical analysis was performed using one-way ANOVA with Prism 9 software (GraphPad). Results are expressed as mean ± standard error (SEM). Dunnett's test was used for post-hoc pairwise comparisons. Values ​​of *P < 0.05, **P < 0.01, ***P < 0.001, or ****P < 0.001 indicate statistical significance.

[0166] References 1-Lebrin, F.; Srun, S.; Raymond, K.; Martin, S.; van den Brink, S.; Freitas, C.; Breant, C.; Mathivet, T.; Larrivee, B.; Thomas, JL; Arthur, HM; Westermann, CJ; Disch, F.; Vessel maturation and reduces epistaxis in individuals with hereditary hemorrhagic telangiectasia. Nat Med 2010, 16, 420-8. 2-Lebrin, FS, C.; Thalgott, J. Use of thalidomide or analogs thereof for preventing neurologic disorders induced by brain irradiation. 2015. 3-Cheng J, Jiang J, He B, Lin WJ, Li Y, Duan J, Li H, Huang X, Cai J, Xie J, Zhang Z, Yang Y, Xu Y, Hu X, Wu M, Zhuo X, Liu Q, Shi Z, Yu P, Rong X, Ye X, Saw PE, Wu LJ, Simone CB 2nd, Chua MLK, Mai HQ, Tang Y. A phase 2 study of thalidomide for the treatment of radiation-induced-blood-brain barrier injury. .Sci Transl Med. 2023 Feb 22;15(684):eabm6543. doi: 10.1126 / scitranslmed.abm6543. Epub 2023 Feb 22.PMID: 36812346 Clinical Trial. 4-Mishra, A.; O'Farrell, F. M.; Reynell, C.; Hamilton, N. B.; Hall, C. N.; Attwell, D. Imaging pericytes and capillary diameter in brain slices and isolated retinae. Nat Protoc 2014, 9, 323-36. 5-Armulik, A.; Genove, G.; Mae, M.; Nisancioglu, M. H.; Wallgard, E.; Niaudet, C.; He, L.; Norlin, J.; Lindblom, P.; Strittmatter, K.; Johansson, B. R.; Betsholtz, C. Pericytes regulate the blood-brain barrier. Nature 2010, 468, 557-61. 6-Mountain, A.; Nikolakopoulou, AM; Zhao, Z.; Sagar, AP; Si, G.; Lazic, D.; Barnes, SR; Daianu, M.; Ramanathan, A.; Go, A.; Lawson, EJ; Wang, Y.; Mack, WJ; Thompson, PM; Schneider, JA; Varkey, J.; Langen, R.; Mullins, E.; Jacobs, RE; Zlokovic, BV Pericyte degeneration causes white matter dysfunction in the mouse central nervous system. Nat Med 2018, 24, 326-337.

Claims

1. A formulation comprising a pharmaceutical compound and a pharmaceutically acceptable additive, adjuvant, diluent and / or carrier, wherein the pharmaceutical compound is of formula I or II: 【Chemistry 1】 [During the ceremony, A １ A ２ A ３ and A ４ Each of them operates independently, CR １ Or it represents N; A ５ and A ６ These are S and CR, which are independent of each other. １ Or it represents N; R １ represents hydrogen, halogen, OH, C １ to C ８ alkyl, NO ２ , or NR ２ R ３ and R ２ and R ３ These are H and C, respectively, independently. １ ~C ８ Alkyl, C １ ~C ８ -Alkyl-carbonyl, and C １ ~C ８ Selected from alkoxy-carbonyl groups; R ｘ is Y or CHR ４ R ５ This represents, Y represents a four-membered, five-membered, or six-membered heterocycle containing one, two, or three nitrogen atoms. Y is optionally a halide, saturated, or unsaturated C １ ~C ８ Hydrocarbons, C １ ~C ８ -alkoxy-carbonyl, or C １ ~C ８ It may be substituted with one or more substituents selected from the group consisting of alkyl-carbonyl, preferably acetyl or propionyl; R ４ H, C may be optionally substituted at one or more carbon atoms with one or more groups selected from halogens, aminos, sulfinyls, and sulfonyls. １ ~C ８ - Alkyl, C １ ~C ８ - Cycloalkyl, C １ ~C ８ - Cycloalkylalkyl, heteroaryl, C １ ~C ８ - Heteroaralkyl, C １ ~C ８ - Heterocycloalkyl, or C １ ~C ８ - Represents heterocycloalkylalkyl, R ５ H, OH, COOH, or C may be substituted at one or more carbon atoms with one or more groups selected from halogens, aminos, sulfinyls, and sulfonyls, preferably COOH, i-propyl, or t-butyl. １ ~C ８ - Represents alkyl; However, R ４ and R ５ [Except when both are H] The preparation having the structure shown in [figure name].

2. A １ Or A ２ C-NH ２ or C-NHC(O)CH ３ It represents, preferably A １ C-NH ２ It represents and A ２ A ３ and A ４ Each of these is independently CH and C-CH. ３ Alternatively, it represents C-Cl, preferably CH, or A ２ C-NH ２ It represents and A １ A ３ and A ４ Each of these is independently CH and C-CH. ３ The formulation according to claim 1, or representing C-Cl, preferably C.

3. A １ However, C-NH ２ It represents and A ２ A ３ and A ４ However, CH and CH are independent of each other. ３ The formulation according to claim 1, or comprising CCl, preferably CH.

4. R ｘ but, 【Chemistry 2】 [In the formula, R ６ C １ ~C ８ - Alkyl, C ２ ~C ８ - Alkenil, C ２ ~C ８ - Alkinyl, C １ ~C ８ - Alkoxyalkyl, C １ ~C ８ -alkoxy-carbonyl, aryl, or C １ ~C ８ - Represents alkyl-carbonyl, preferably acetyl or propionyl. A formulation according to claim 1 or 2, which represents the following:

5. R ｘ However, CHR ４ R ５ This represents, and here, R ４ The formulation according to claims 1 to 3, wherein is represented by hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, tert-butyl, neopentyl, isopentyl, cyclopropyl-methyl, or thienyl-methyl.

6. R ｘ represents CHR ４ R ５ where R ５ represents COOH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, tert-butyl, preferably COOH, methyl, i-propyl or t-butyl, the preparation according to any one of claims 1 to 3 or 5.

7. The pharmaceutical compound is of formula I-a: 【Transformation 3】 [wherein each Z is independently hydrogen, halogen, OH, C １ to C ８ alkyl, NO ２ , and NR ２ R ３ selected from, R ２ and R ３ are each independently H, C １ to C ８ alkyl, C １ to C ８ -alkyl-carbonyl, and C １ to C ８ -alkoxy-carbonyl selected from, preferably, each Z is independently H, halogen, NH ２ , and NH C(O)CH ３ selected from; m is 1, 2, 3, or 4, preferably 1 or 2; n is 1, 2, or 3, and R ６ is saturated or unsaturated C １ ~C ８ Hydrocarbons, C １ ~C ８ -alkoxy-carbonyl, or C １ ~C ８ - Represents alkyl-carbonyl, preferably acetyl or propionyl. The formulation according to claim 1, having the structure shown in [image / figure].

8. The pharmaceutical compound is of formula I-b or I-c: 【Chemistry 4】 [In the formula, each Z is independently hydrogen, halogen, OH, C] １ ~C ８ Alkyl, and NO ２ Selected from, preferably, each Z is independently H, CH ３ and selected from Cl; m is 1, 2, or 3; and R ６ is saturated or unsaturated C １ ~C ８ Hydrocarbons, C １ ~C ８ -alkoxy-carbonyl, or C １ ~C ８ - Represents alkyl-carbonyl, preferably acetyl or propionyl. The formulation according to claim 1, having the structure shown in [image / figure].

9. The formulation according to any one of claims 1 to 8, wherein the pharmaceutical compound does not bind to cereblon (CRBN).

10. The pharmaceutical compound, 【Transformation 5】 【Transformation 6】 【Transformation 7】 A formulation according to any one of claims 1 to 9, selected from the above.

11. Formula I or II: 【Transformation 8】 [During the ceremony, A １ A ２ A ３ and A ４ Each of them operates independently, CR １ Or it represents N; A ５ and A ６ These are S and CR, which are independent of each other. １ Or it represents N; R １ is hydrogen, halogen, OH, C １ ~C ８ Alkyl, NO ２ , or NR ２ R ３ This represents, R ２ and R ３ These are H and C, respectively, independently. １ ~C ８ Alkyl, C １ ~C ８ -Alkyl-carbonyl, and C １ ~C ８ Selected from alkoxy-carbonyl groups; R ｘ CHR ４ R ５ This represents, R ４ H, C １ ~C ８ - Alkyl, C １ ~C ８ - Cycloalkyl, C １ ~C ８ - Cycloalkylalkyl, heteroaryl, C １ ~C ８ - Heteroaralkyl, C １ ~C ８ - Heterocycloalkyl, or C １ ~C ８ - Represents heterocycloalkylalkyl, preferably hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, tert-butyl, neopentyl, isopentyl, cyclopropyl-methyl, or thienyl-methyl; and R ５ is H, OH, COOH, or C １ ~C ８ Alkyl, preferably OH, COOH, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, or tert-butyl, preferably COOH, methyl, i-propyl, or t-butyl, However, R ４ and R ５ [Except when both are H] A pharmaceutical compound having the structure shown.

12. Formula I or II for medicinal use in the treatment of diseases or disorders characterized by pericyte dysfunction, pericyte detachment, and / or pericyte loss: 【Chemistry 9】 [During the ceremony, A １ A ２ A ３ and A ４ Each of them operates independently, CR １ Or it represents N; A ５ and A ６ These are S and CR, which are independent of each other. １ Or it represents N; R １ is hydrogen, halogen, OH, C １ ~C ８ Alkyl, NO ２ , or NR ２ R ３ This represents, R ２ and R ３ These are H and C, respectively, independently. １ ~C ８ Alkyl, C １ ~C ８ -Alkyl-carbonyl, and C １ ~C ８ Selected from alkoxy-carbonyl groups; R ｘ is H, Y, or CHR ４ R ５ This represents, Y represents a four-membered, five-membered, or six-membered heteroring containing one, two, or three heteroatoms. Y is optionally a halide, saturated, or unsaturated C １ ~C ８ Hydrocarbons, C １ ~C ８ -alkoxy-carbonyl, or C １ ~C ８ It may be substituted with one or more substituents selected from the group consisting of alkyl-carbonyl, preferably acetyl or propionyl; R ４ H, C １ ~C ８ - Alkyl, C １ ~C ８ - Cycloalkyl, C １ ~C ８ - Cycloalkylalkyl, heteroaryl, C １ ~C ８ - Heteroaralkyl, C １ ~C ８ - Heterocycloalkyl, or C １ ~C ８ - Represents heterocycloalkylalkyl, R ５ is H, OH, COOH, or C １ ~C ８ [Represents alkyl, preferably COOH, i-propyl, or t-butyl] A pharmaceutical compound having the structure shown, or a formulation comprising such a compound with a pharmaceutically acceptable additive, adjuvant, diluent and / or carrier.

13. A compound or formulation for use according to claim 12, wherein the disease or disorder characterized by pericyte dysfunction, detachment and / or loss is selected from diabetic complications, chronic kidney disease (CKD), small vessel disease, and central nervous system (CNS) diseases.

14. A compound or formulation for use according to claim 12, wherein the disease or disorder characterized by pericyte dysfunction, detachment and / or loss is selected from diabetic nephropathy (DN), diabetic retinopathy (DR), diabetic cardiomyopathy (DCM), and diabetic neuropathy.

15. The compound or formulation for use according to claim 12, wherein the disease or disorder characterized by pericyte dysfunction, detachment and / or loss is selected from hypertensive nephropathy, IgA nephropathy, congenital nephrotic syndrome, lupus nephritis, polycystic kidney disease and allograft nephropathy.

16. A compound or formulation for use according to claim 12, wherein the disease or disorder characterized by pericyte dysfunction, detachment and / or loss is selected from autosomal dominant cerebral arteriopathies with subcortical infarction and leukoencephalopathy (Cadasil), autosomal recessive cerebral arteriopathies with subcortical infarction and leukoencephalopathy (Carasil), cerebral amyloid angiopathy (CAA), retinal vascular disorder with cerebral leukoencephalopathy and systemic symptoms (RVCL-S), hereditary hemorrhagic telangiectasia (HHT), and cavernous hemangioma (CCM).

17. A compound or formulation for use according to claim 12, wherein the disease or disorder characterized by pericyte dysfunction, detachment and / or loss is selected from stroke, epilepsy, spinal cord injury, vascular dementia, Alzheimer's disease, Huntington's disease, Parkinson's disease, traumatic brain injury, multiple sclerosis, amyotrophic lateral sclerosis, and radiation necrosis.

18. A pharmaceutical formulation according to any one of claims 1 to 10, for use as a pharmaceutical in treatments for vascular stabilization and / or treatments for promoting, restoring, or maintaining pericyte adhesion to blood vessels.

19. A formulation for use according to claim 18, wherein the therapeutic procedure for vascular stabilization includes protecting the integrity of the blood-brain barrier, controlling blood flow, limiting inflammation and / or reducing tissue damage.

20. A method for promoting, restoring, or maintaining pericyte attachment to blood vessels in a patient, the method comprising administering to the patient a formulation according to any one of claims 1 to 10.

21. A method for treating a vascular disease in a patient, the method comprising administering to the patient a formulation according to any one of claims 1 to 10.

22. Vascular diseases include diabetic complications, chronic kidney disease (CKD), small vessel diseases, and central nervous system (CNS) diseases; preferably hypertensive nephropathy, IgA nephropathy, congenital nephrotic syndrome, lupus nephritis, polycystic kidney disease, and allograft nephropathy; autosomal dominant cerebral arteriovenous disease (Cadasil) with subcortical infarction and leukoencephalopathy, autosomal recessive cerebral arteriovenous disease (Carasil) with subcortical infarction and leukoencephalopathy, and cerebral amyloidosis. The method according to claim 21, selected from dangiopathies (CAA), retinal vascular disorders with cerebral leukoencephalopathy and systemic symptoms (RVCL-S), hereditary hemorrhagic telangiectasia (HHT), and cavernous hemangioma (CCM); stroke, epilepsy, spinal cord injury, vascular dementia, Alzheimer's disease, Huntington's disease, Parkinson's disease, traumatic brain injury, multiple sclerosis, amyotrophic lateral sclerosis, and radiation necrosis.

23. A method for maturing an organoid and / or organ-on-a-chip model system, comprising using a formulation according to any one of claims 1 to 10 or a compound defined in any one of claims 1 to 10.

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