Novel pomalidomide derivatives, their production method and use

Abstract

The present invention relates to a novel pomalidomide derivative that exhibits improved effects compared to pomalidomide, and its manufacturing method and use. The novel pomalidomide derivative according to the present invention has excellent binding ability to cereblon, almost no cytotoxicity, low possibility of teratogenic side effects, can regulate TNF-α and proinflammatory cytokine expression, and has excellent pharmacokinetic stability in vivo while suppressing oxidative stress, and is expected to have improved effects compared to commercially available thalidomide drugs.

Description

[Technical field]

[0001] The present invention relates to novel pomalidomide derivatives that exhibit improved pharmacological effects compared to pomalidomide, and to a method for producing the same and use thereof. [Background technology]

[0002] Thalidomide was a drug sold in the late 1950s and 1960s to prevent morning sickness in pregnant women, but due to reports of teratogenic side effects, it was banned for use by pregnant women, as well as those of childbearing age and those who may become pregnant. However, interest in the drug has been rekindled after thalidomide was found to be clinically effective in treating erythema nodosum (ENL), HIV wasting syndrome, and various cancers.

[0003] Mechanistic studies on ENL activity confirmed an anti-tumor necrosis factor alpha (anti-TNF-α) effect; specifically, thalidomide increases the degradation of TNF-α RNA and decreases its synthesis and secretion. Further studies have shown that it is a known costimulator of both CD8+ and CD4+ T cells, an inhibitor of basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and the transcription factor NF-κB, and can also act as an angiogenesis inhibitor.

[0004] TNF-α and its family members play pivotal roles in various physiological and pathological processes, including cell proliferation and differentiation, cell death, regulation of immune responses, and induction of inflammation. TNF-α acts through two receptors, TNFR1 and TNFR2, the former of which is expressed in all tissues and is the main signaling receptor for TNF-α. The latter is expressed mainly in immune cells and mediates more restricted biological responses. When cells are exposed to TNF-α, the caspase cascade is activated, which can lead to the death of the cell through cell apoptosis. Indeed, the main cell surface molecules that can initiate cell death are the ligands and receptors of the TNF family members. For example, each death-inducing member of the TNF receptor family contains a cytoplasmic "death domain" (DD), a protein-protein interaction motif important for binding to downstream components of the signaling machinery.

[0005] Recently, it has been shown that tumor necrosis factor-related apoptosis-inducing ligand TRAIL selectively induces cell death in tumor cells, but not in most normal cells. TRAIL is known to mediate cell death in thymocytes and play an important role in the induction of autoimmune diseases. More frequently, however, TNF-α receptor binding induces the activation of transcription factors, AP-1 and NF-κB, which then induce genes involved in acute and chronic inflammatory responses. Thus, overproduction of TNF-α is involved in many inflammatory diseases such as rheumatoid arthritis, graft-versus-host disease and Crohn's disease, and is known to further exacerbate ENL, septic shock, and dementia associated with AIDS and Alzheimer's disease (AD).

[0006] Thalidomide-based therapeutic agents available on the market include thalidomide, bortezomib, lenalidomide, and pomalidomide. Injectable bortezomib is steadily maintaining its market share and showing a growing trend, while oral lenalidomide has replaced the position of existing thalidomide and is showing a high growth trend in the market. Lenalidomide is the next generation drug of thalidomide, and shows a more excellent therapeutic effect than thalidomide due to its stronger cancer cell killing and immune regulation. It is known that the combination of lenalidomide and dexamethasone for treatment in cases of recurrence or refractory disease with existing treatment is very effective, with a disease-free survival of 13.4 months and an overall survival of 38 months. As for side effects, problems such as peripheral neuropathy that were seen with existing thalidomide have almost disappeared, and although the bone marrow suppression side effects have become slightly worse, it is known that there are no major problems if leukocyte-stimulating factors are administered. Pomalidomide was approved by the US FDA in 2013 for the treatment of relapsed and refractory multiple myeloma, and is used in people who have received at least two conventional therapies, including lenalidomide and bortezomib, and whose disease progresses within 60 days of completing the last treatment. Pomalidomide directly inhibits angiogenesis and myeloma cell growth, but this dual effect is centered on its activity in myeloma rather than other pathways such as suppression of TNF-α. Pomalidomide exhibits antiangiogenic and antimyeloma activity through suppression of IL-6 expression as well as enhancement of expression of IFN-γ, IL-2, and IL-10.

[0007] In U.S. Patent No. 9,623,020, it was discovered that new thio compounds were synthesized using thalidomide derivatives, and that these thio compounds could regulate TNF-α activity and change the expression of immune cytokines by TCR-stimulated T cells.

[0008] The inventors have made extensive efforts to develop further derivatives to improve the pharmaceutical effects of thalidomide, and have confirmed that the compound of [Chemical Formula 1] exhibits pharmaceutical effects equal to or greater than those of conventional thalidomide derivatives and has significantly improved pharmacokinetic stability in vivo, thereby completing the present invention. Summary of the Invention [Problem to be solved by the invention]

[0009] An object of the present invention is to provide a novel thalidomide derivative which exhibits improved pharmacological effects as compared with conventionally known thalidomide drugs, a method for producing the same, and uses thereof. [Means for solving the problem]

[0010] In order to achieve the above object, the present invention provides a compound represented by [Chemical Formula 1]:

[0011] [ka]

[0012] The present invention also provides a method for preparing the compound, comprising: (a) dissolving 3,6'-dithiopomalidomide in a mixed solvent of DMSO and H2O, and heating the mixture.

[0013] In the present invention, the mixed solvent in step (a) is characterized in that DMSO:H2O is mixed in a volume ratio of 1:0.1-5.

[0014] In the present invention, the heating in the step (a) is characterized by heating at 80 to 120° C. for 5 to 30 hours.

[0015] In the present invention, the method further comprises, after step (a), a step (b) of purifying the compound.

[0016] In the present invention, the purification in step (b) is characterized in that the solution produced in step (a) is diluted with EtOAc, the organic layer is dried, and the organic layer is purified by SiO2 column chromatography.

[0017] The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease, which comprises the compound or a pharma- ceutically acceptable salt thereof as an active ingredient.

[0018] In the present invention, the inflammatory disease is characterized in that it is selected from the group consisting of psoriasis, rheumatoid arthritis and Crohn's disease.

[0019] The present invention also provides a method for preventing or treating an inflammatory disease, comprising administering a compound of [Chemical Formula 1] or a pharma- ceutically acceptable salt thereof, or a composition containing the compound of [Chemical Formula 1] or a pharma- ceutically acceptable salt thereof, to a subject in need thereof.

[0020] [ka]

[0021] The present invention also provides a use of a compound of formula 1 or a pharma- ceutically acceptable salt thereof for the prophylaxis or treatment of an inflammatory disease:

[0022] [ka]

[0023] The present invention also provides a use of a compound of formula 1 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for the prophylaxis or treatment of an inflammatory disease:

[0024] [ka]

[0025] The present invention also provides a pharmaceutical composition for preventing or treating angiogenic diseases, which comprises the compound or a pharma- ceutically acceptable salt thereof as an active ingredient.

[0026] In the present invention, the angiogenic disease is characterized in that it is selected from the group consisting of corneal transplant angiogenesis, neovascular glaucoma, diabetic retinopathy, exudative age-related macular degeneration, diabetic macular edema, corneal diseases caused by neovascularization, macular degeneration, pterygium, retinal degeneration, retrolental fibroplasia, granular conjunctivitis, hemangioma, angiofibroma, vascular malformation, arteriosclerosis, vascular adhesion and edematous sclerosis.

[0027] The present invention also provides a method for preventing or treating angiogenic diseases, comprising administering a compound of [Chemical Formula 1] or a pharma- ceutical acceptable salt thereof, or a composition containing the compound as an active ingredient, to a subject in need thereof.

[0028] [ka]

[0029] The present invention also provides a use of a compound represented by formula 1 or a pharma- ceutically acceptable salt thereof for the prophylaxis or treatment of angiogenic diseases:

[0030] [ka]

[0031] The present invention also provides a use of a compound of formula 1 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of angiogenic diseases:

[0032] [ka] Effect of the Invention

[0033] The novel pomalidomide derivative developed in this invention has excellent binding affinity to cereblon, is almost non-cytotoxic, has a low possibility of teratogenic side effects, and can regulate TNF-α and pro-inflammatory cytokine expression. It also has excellent pharmacokinetic stability in the body while suppressing oxidative stress, and is expected to have improved effects compared to commercially available thalidomide-based drugs. [Brief description of the drawings]

[0034] [Figure 1] Figure 1 shows the cereblon-binding activity of 3-MP compared to that of pomalidomide through in vitro competitive inhibition reactions. [Diagram 2] Figure 2 shows the results of evaluating the cytotoxicity of pomalidomide and 3-MP against the MM.1S cell line. [Figure 3a] Figure 3a shows the results of immunoblotting to compare the intracellular Ikaros and Aiolos degradation effects when pomalidomide and 3-MP were treated in MM.1S cells. [Figure 3b] Figure 3b shows the results of immunoblotting to compare the decomposition effects of intracellular Ikaros and Aiolos at different times when MM.1S cells were treated with pomalidomide and 3-MP, respectively. [Figure 4] FIG. 4 shows the results of immunoblotting to compare the extent of intracellular SALL4 degradation in Tera-1 cells treated with pomalidomide and 3-MP. [Figure 5a] Figure 5a shows the results of RT-PCR comparing the inhibitory effects of pomalidomide and 3-MP on the production of pro-inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-8 at the mRNA level when PBMC cells were treated with the drugs. [Figure 5b] Figure 5b shows the results of RT-PCR comparing the inhibitory effects of pomalidomide and 3-MP on the production of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 at the mRNA level when BV-2 cells were treated with the drugs. [Figure 6a]Figure 6a shows the results of treating RAW264.7 cells with pomalidomide and 3-MP, respectively, and comparing the oxidative stress inhibitory effects by immunoblotting for iNOS and COX-2. [Figure 6b] Figure 6b shows the results of treating BV-2 cells with pomalidomide and 3-MP, respectively, and comparing the oxidative stress inhibitory effects by measuring the nitrite content in the cell culture medium. [Figure 6c] Figure 6c shows the results of treating BV-2 cells with pomalidomide and 3-MP, respectively, and comparing the oxidative stress inhibitory effects by immunoblotting for iNOS and COX-2. [Figure 6d] Figure 6d shows the results of RT-PCR comparing the oxidative stress inhibitory effects of pomalidomide and 3-MP on BV-2 cells at the mRNA levels of iNOS and COX-2. [Figure 7a] Figure 7a shows the calibration curve for the plasma stability analysis of 3-MP. The linear equation of the calibration curve and the correlation coefficient (r) are displayed on the graph. The Y-axis shows the peak area of ​​the standard and the X-axis shows the concentration (μg / mL). [Figure 7b] FIG. 7b shows the results of analyzing the stability of 3-MP in human plasma over time. BEST MODE FOR CARRYING OUT THEINVENTION

[0035] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by a skilled artisan in the art to which this invention belongs. Generally, the nomenclature used herein is one that is well known and commonly used in the art.

[0036] In the present invention, a thio compound was prepared by improving pomalidomide, and the pharmaceutical efficacy of the novel compound was confirmed.

[0037] Specifically, it was confirmed that the novel compound according to the present invention has superior cereblon binding activity and less cytotoxicity compared to its parent compound, pomalidomide, and is therefore superior in in vivo safety.In addition, it was found that the novel compound according to the present invention does not induce the degradation of cereblon substrates Aiolos, Ikaros, and SALL4 as strongly as pomalidomide, thereby reducing the main side effects caused by thalidomide-based compounds.

[0038] Furthermore, it has been revealed that the novel compound according to the present invention can suppress the expression of various pro-inflammatory cytokines, suppress oxidative stress, and maintain in vivo stability for a long period of time via various administration routes, such as intravenous administration, oral administration, and intraperitoneal administration.

[0039] Thus, in one aspect, the present invention relates to novel compounds represented by formula 1:

[0040] [ka]

[0041] In the present invention, the compound of [Chemical Formula 1] can be named 3-thiopomalidomide, C 13 H 13 It can be represented by the chemical formula N3O3S and has a molecular weight of 289.31. In the present invention, the compound of [Chemical Formula 1] can be abbreviated as 3-MP.

[0042] In another aspect, the present invention relates to a method for preparing a compound of formula (1), comprising the steps of: (a) dissolving 3,6'-dithiopomalidomide in a mixed solvent of DMSO and HO; and heating the mixture.

[0043] In the present invention, the mixed solvent in step (a) is characterized in that DMSO:HO is mixed in a volume ratio of 1:0.1-5, preferably 1:0.2-1, and more preferably 1:0.3-0.8, but is not limited thereto.

[0044] In the present invention, the step (a) is characterized by dissolving 0.1 to 20 mmol of 3,6'-dithiopomalidomide per 100 ml of the mixed solvent, preferably 1 to 10 mmol of 3,6'-dithiopomalidomide per 100 ml of the mixed solvent, more preferably 3 to 5 mmol of 3,6'-dithiopomalidomide per 100 ml of the mixed solvent, but is not limited thereto.

[0045] In the present invention, the heating in step (a) is characterized by heating at 80 to 120°C for 5 to 30 hours, the temperature range is preferably 90 to 110°C, more preferably 95 to 105°C, and the heating time is preferably 10 to 25 hours, more preferably 15 to 20 hours, but is not limited thereto.

[0046] In the present invention, the method further comprises, after the heating in step (a), a step (b) of purifying the compound.

[0047] In the present invention, the purification in step (b) is characterized by diluting the solution produced in step (a) with EtOAc, drying the organic layer, and purifying it by SiO2 column chromatography, but is not limited thereto.

[0048] In the present invention, the solution diluted with EtOAc may be washed, and the washing is characterized by being washed with H2O and / or salt water, but is not limited thereto. The washing is preferably washed with H2O twice and salt water once or more.

[0049] In the present invention, after the washing, EtOAc can be added to extract the aqueous layer.

[0050] In the present invention, after the aqueous layer is extracted, the organic layers can be combined and dried, which is characterized by, but not limited to, drying over MgSO4.

[0051] In the present invention, the dried product can be concentrated after filtration, and the residue can be purified by column chromatography to increase the purity of the compound of the present invention.

[0052] In the present invention, the column chromatography is characterized in that it is silica gel (SiO2) column chromatography, and preferably, amino silica gel (NH2-SiO2) or reverse phase silica gel (C 18 The method is characterized by, but not limited to, column chromatography (-SiO2).

[0053] When purifying using SiO2 column chromatography, the column chromatography is characterized in that 1-2 g of the residue is dissolved in a mixture of acetone and 1-5 mL of DMSO, coated, mixed with silica gel, and then the silica gel that has been rotary evaporated is loaded, and then the compound of the present invention is separated using a solution mixed with CHCl3:acetone = 20:1-9:1, but is not limited thereto.

[0054] In the present invention, the compound of the present invention can be further purified through recrystallization, in which case the solvent used in the recrystallization method is any one or more selected from the group consisting of CHCl3, CH2Cl2 and EtOAc, but is not limited thereto.

[0055] In another aspect, the present invention relates to a pharmaceutical composition for preventing or treating an inflammatory disease, which contains the above-mentioned compound as an active ingredient.

[0056] In the present invention, the inflammatory disease is characterized in that it is selected from the group consisting of psoriasis, rheumatoid arthritis and Crohn's disease, but is not limited thereto.

[0057] In another embodiment, the present invention relates to a method for preventing or treating an inflammatory disease, comprising administering to a subject in need thereof a compound represented by [Chemical Formula 1] or a pharma- ceutically acceptable salt thereof, or a composition containing the compound as an active ingredient:

[0058] [ka]

[0059] In yet another aspect, the present invention also relates to a use of a compound of [Chemical Formula 1] or a pharma- ceutically acceptable salt thereof for the prophylaxis or treatment of an inflammatory disease:

[0060] [ka]

[0061] In yet another aspect, the present invention provides a use of a compound of formula 1 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for the prophylaxis or treatment of an inflammatory disease:

[0062] [ka]

[0063] In another aspect, the present invention relates to a pharmaceutical composition for preventing or treating angiogenic diseases, which contains the above-mentioned compound as an active ingredient.

[0064] In the present invention, the angiogenic disease is characterized by being selected from the group consisting of corneal transplant angiogenesis, neovascular glaucoma, diabetic retinopathy, exudative age-related macular degeneration, diabetic macular edema, corneal diseases caused by neovascularization, macular degeneration, pterygium, retinal degeneration, retrolental fibroplasia, granular conjunctivitis, hemangioma, angiofibroma, vascular malformation, arteriosclerosis, vascular adhesion and edematous sclerosis, but is not limited thereto.

[0065] In another embodiment, the present invention relates to a method for preventing or treating angiogenic diseases, comprising administering to a subject in need thereof a compound represented by [Chemical Formula 1] or a pharma- ceutically acceptable salt thereof, or a composition containing the compound as an active ingredient:

[0066] [ka]

[0067] In yet another aspect, the present invention also relates to a use of a compound of [Chemical Formula 1] or a pharma- ceutically acceptable salt thereof for the prophylaxis or treatment of angiogenic diseases:

[0068] [ka]

[0069] In yet another aspect, the present invention provides a use of a compound of formula 1 or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for the prophylaxis or treatment of angiogenic diseases:

[0070] [ka]

[0071] In the present invention, the term "pharmaceutical composition" or "pharmaceutical formulation" refers to a mixture containing a pharma- ceutical acceptable excipient, such as a diluent or carrier, that renders the novel compound of the present invention particularly suitable for in vivo or in vitro diagnostic or therapeutic use. According to some embodiments, a pharmaceutical composition containing the composition of the present invention can be provided in a manner that allows it to be administered in a therapeutically effective amount to a subject in need thereof. In some embodiments, the composition of the present invention can be administered to a human.

[0072] As used herein, an "effective amount" or "therapeutically-effective amount" refers to an amount of a compound or composition (e.g., a compound or composition of the invention) sufficient to achieve a beneficial or desired result. An effective amount can be administered in one or more administrations, applications or dosages, and is not intended to be limited to a particular formulation or route of administration.

[0073] In the present invention, the term "pharmaceutically acceptable salt" refers to an acid that forms a non-toxic acid addition salt containing a pharmaceutically acceptable anion, for example, an acid addition salt formed by inorganic acids such as sulfuric acid, hydrochloric acid, nitric acid, phosphoric acid, hydrobromic acid, hydroiodic acid, etc.; organic acids such as tartaric acid, formic acid, citric acid, acetic acid, trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, lactic acid, malonic acid, malic acid, salicylic acid, succinic acid, oxalic acid, propionic acid, aspartic acid, glutamic acid, citric acid, etc.; and sulfonic acids such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, etc. The compounds represented by [Chemical Formula 1] according to the present invention, which include free carboxy substituents, may be in the form of salts, such as the above-mentioned acid addition salts and sodium, calcium and ammonium salts, and may also be pharma- ceutically acceptable salts, such as alkali metal or alkaline earth metal salts formed with lithium, sodium, potassium, calcium, magnesium, etc., amino acid salts such as lysine, arginine, guanidine, etc., and organic salts such as dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, diethanolamine, choline, triethylamine, etc.

[0074] The compound of Formula 1 according to the present invention can be converted into its salt by a conventional method, and the preparation of the salt can be easily performed by those skilled in the art based on the structure of Formula 1 without any special description.

[0075] Although the pharmaceutical composition provided in the present invention is primarily intended for administration to humans, a person skilled in the art can understand that such compositions are generally suitable for administration to all kinds of animals. That is, the pharmaceutical composition according to the present invention can also be administered to animals requiring veterinary treatment, such as other mammals, such as livestock (e.g., dogs, cats, etc.), farm animals (e.g., cows, sheep, pigs, horses, etc.) and laboratory animals (e.g., rats, mice, guinea pigs, etc.). A skilled veterinary pharmacologist who is familiar with the modifications of pharmaceutical compositions for administration to various animals can design and / or perform such modifications simply by routine experimentation, if necessary.

[0076] The pharmaceutical compositions described in this invention may be prepared by any method known in the pharmacological art or as described below. In general, such methods for preparation include the step of bringing the active ingredient into association with an excipient and / or one or more other accessory ingredients and then, if necessary or desired, shaping and / or packaging the product into the desired single or multi-dose unit.

[0077] The pharmaceutical composition of the present invention can be manufactured, packaged, and / or sold unpackaged in a single unit dose and / or in a plurality of single unit doses.The term "unit dose" as used herein refers to a discrete amount of pharmaceutical composition that contains a predetermined amount of active ingredient.The amount of active ingredient is generally the same as the dosage of active ingredient administered to a subject and / or a convenient fraction of such a dosage, for example, 1 / 2 or 1 / 3 of the dosage.

[0078] The relative amounts of active ingredient, pharma- ceutically acceptable excipient, and / or any additional ingredients in a pharmaceutical composition of the invention will vary depending on the identity, size, and / or condition of the subject being treated, and on the route by which the composition is administered. By way of example, the composition may contain from 0.001% to 100% (w / w) active ingredient.

[0079] As used in the present invention, pharma- ceutically acceptable excipients include any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, etc., suitable for a particular dosage form purpose. Remington's The Science and Practice of Pharmacy, 21st Edition, AR Gennaro, (Lippincott, Williams & Wilkins, Baltimore, MD, 2006) discloses various excipients used in the preparation of pharmaceutical compositions and known techniques for their manufacture. Any common carrier medium is considered within the scope of the present invention, except that it is incompatible with the substance or its derivatives, for example by providing any undesirable biological effect or interacting in a deleterious manner with any other component of the pharmaceutical composition. Pharmaceutically acceptable excipients are at least 95%, 96%, 97%, 98%, 99%, or 100% pure.

[0080] The excipients are approved for human and veterinary use. In some embodiments, the excipients are approved by the U.S. Food and Drug Administration. In some embodiments, the excipients are pharmaceutical grade. In some embodiments, the excipients meet the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and / or the International Pharmacopoeia (EP).

[0081] Pharmaceutically acceptable excipients used in the preparation of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surfactants and / or emulsifying agents, disintegrants, binders, preservatives, buffers, lubricants, and / or oils.

[0082] Such excipients can optionally be included in the formulations of the present invention. Excipients, such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening, flavoring, and perfuming agents, can be present in the composition, according to the discretion of the formulator.

[0083] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, calcium hydrogen phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and combinations thereof.

[0084] Exemplary granulating and / or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clay, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood, natural sponge, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked sodium carboxymethylcellulose (croscarmellose), methylcellulose, pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and combinations thereof.

[0085] Exemplary surfactants and / or emulsifiers include natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminum silicate)), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, ethylene glycol distearate, glyceryl monostearate, and propylene glycol monostearate, polyvinyl alcohol), carbomers (e.g., carboxypolymethylene, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers), carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan, sorbitol, sorbitol derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitol, ... fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60), polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate (Span 40), sorbitan monostearate (Span 60), sorbitan tristearate (Span 65), glyceryl monooleate, sorbitan monooleate (Span 80)), polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj45), ... Examples of suitable oleic acid salts include ethylene glycol hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Solutol), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether (Brij30)), poly(vinylpyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate,Including, but not limited to, Pluronic F-68, Poloxamer P-188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and / or combinations thereof.

[0086] Exemplary binders include, but are not limited to, starches (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol, and the like), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husk, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, poly(vinylpyrrolidone), magnesium aluminum silicate (Veegum), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and combinations thereof.

[0087] Exemplary preservatives may include antioxidants, chelating agents, antibacterial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives. Exemplary anti-depressants include, but are not limited to, alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and trisodium edetate. Exemplary antimicrobial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal. Exemplary antifungal preservatives include, but are not limited to, butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid. Exemplary alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol. Exemplary acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.Other preservatives include, but are not limited to, tocopherol, tocopherol acetate, deteroxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, and Euxyl. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0088] Exemplary buffering agents include, but are not limited to, citrate buffer solution, acetate buffer solution, phosphate buffer solution, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasic calcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dibasic potassium phosphate, monobasic potassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, dibasic sodium phosphate, monobasic sodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and combinations thereof.

[0089] Exemplary lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behenate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and combinations thereof.

[0090] Exemplary natural oils include almond, apricot kernel, avocado, babassu, bergamot, black current seed, borage, cade, chamomile, canola, caraway, carnauba, castor oil, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cottonseed, emu, eucalyptus, evening primrose, fish, linseed, geraniol, loofah, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, kukui nut, lavandin, lavender, lemon, ritzel, and the like. Exemplary oils include, but are not limited to, sorbitol, macadamia nuts, mallow, mango seeds, meadowfoam seeds, mink, nutmeg, olive, orange, orange roughy, palm, oil palm seeds, peach kernels, peanuts, poppy seeds, pumpkin seeds, rapeseed, rice bran, rosemary, safflower, sandalwood, camellia, savory, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, camellia, vetiver, walnut, and wheat germ. Exemplary oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and combinations thereof.

[0091] Liquid dosage forms for oral and parenteral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs.In addition to active ingredients, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers (e.g., ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, etc.), oils (e.g., cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof.Apart from inert diluents, oral compositions may also contain adjuvants (e.g., wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, and perfumes, etc.). In certain embodiments for parenteral administration, the novel compounds of the present invention are mixed with a solubilizing agent such as Cremophor, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and combinations thereof.

[0092] Injectable preparations, for example, sterile injectable aqueous or oily suspensions, can also be prepared by using dispersing or wetting agents and suspending agents according to known technical methods. Sterile injectable preparations can be sterile injectable solutions, suspensions or emulsions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that can be selected are water, Ringer's solution, USP and isotonic sodium chloride solution. Sterile, fixed oils are also usually selected as solvents or suspending media. For this purpose, any non-irritating fixed oils can be selected, including synthetic mono- or diglycerides. Also, fatty acids, such as oleic acid, are used in the preparation of injectables.

[0093] The injectable preparations can be sterilized, for example, by filtration through a bacterial filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0094] In order to prolong the effect of a drug, it is usually preferable to delay the absorption of the drug from subcutaneous or intramuscular injection.This is accomplished by using a liquid suspension of crystalline or amorphous material with low water solubility.The rate of absorption of the drug is thus dependent on the dissolution rate, which in turn depends on the crystal size and crystalline form.As an alternative, delayed absorption of parenterally administered drugs is achieved by dissolving or suspending the drug in an oil vehicle.

[0095] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharma- ceutically acceptable excipient or carrier (e.g., sodium citrate or dicalcium phosphate), and / or (a) a filler or extender (e.g., starch, lactose, sucrose, glucose, mannitol, and silicic acid), (b) a binder (e.g., carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia), (c) a humectant (e.g., glycerol), (d) a disintegrant (e.g., agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate), (e) a dissolution retarder (e.g., paraffin), (f) an absorption enhancer (e.g., glycerol), (g) a sucrose-containing sorbent (e.g., glycerol), (h) a sucrose-containing sorbent (e.g., glycerol), (i) a sucrose-containing sorbent (e.g., glycerol), (j) a sucrose-containing sorbent (e.g., glycerol), (k) a sucrose-containing sorbent (e.g., sucrose), (m) a sucrose-containing sorbent (e.g., sucrose), (n ... (e.g., quaternary ammonium compounds, etc.), (g) wetting agents (e.g., cetyl alcohol and glycerol monostearate, etc.), (h) absorbents (e.g., kaolin and bentonite clay, etc.), and (i) lubricants (e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, etc.), and mixtures thereof.

[0096] In the case of capsules, tablets, and pills, the dosage form may contain buffering agents. Similar types of solid compositions may be employed as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols. The solid dosage forms of tablets, dragees, capsules, pills, and granules may be prepared with coatings and shells, such as enteric coatings and other coatings well known in the art of pharmacology. They may optionally contain opacifying agents and may be of a composition that they release the active ingredient only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of encapsulating compositions that may be used include polymeric substances and waxes. Similar types of solid compositions may be employed as fillers in soft and hard filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols.

[0097] The active ingredient can be in microencapsulated form with one or more excipients as mentioned above. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings, release-controlling coatings, and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active ingredient can be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Such solid dosage forms can include additional substances other than inert diluents, such as tableting lubricants and other tableting aids, according to conventional methods, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms can include buffering agents. They can optionally include opacifying agents, and can be of a composition such that they release the active ingredient only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of encapsulating agents that can be used include polymeric substances and waxes.

[0098] Dosage forms for topical and / or transdermal administration of the novel compounds of the present invention or pharmaceutical compositions containing the same may include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants and / or patches. Typically, the active ingredient is mixed under sterile conditions with a pharma- ceutical acceptable carrier and / or any necessary preservatives and / or buffers that may be required. In addition, the present invention contemplates the use of transdermal patches, which usually have the added advantage of providing a controlled delivery of the active ingredient to the body. Such dosage forms may be prepared, for example, by dissolving and / or dispersing the active ingredient in a suitable medium. Alternatively or additionally, the rate may be controlled by providing / providing a rate-controlling membrane or dispersing the active ingredient in a polymer matrix and / or gel.

[0099] Formulations for topical administration include, but are not limited to, liquid and / or semi-liquid formulations, such as salves, lotions, oil-in-water and / or water-in-oil emulsions, such as creams, ointments and / or pastes, and / or solutions and / or suspensions. Topically administrable formulations may contain, for example, about 1% to about 10% (w / w) active ingredient, although the concentration of the active ingredient may be as high as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further include one or more additional ingredients described herein.

[0100] The novel compound of the present invention described in this application or pharmaceutical compositions containing it are typically prepared in dosage unit form for easy administration and uniformity of administration. However, it will be understood that the daily usage of the composition of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend on various factors, including the disease, disorder, or disorder being treated and the severity of the disorder, the activity of the specific active ingredient adopted, the specific composition adopted, the subject's age, weight, general health, sex and diet, the administration time, administration route and excretion rate of the specific active ingredient adopted, the duration of treatment, the drugs combined or used simultaneously with the specific active ingredient adopted, and factors well known in the medical field.

[0101] The novel compound of the present invention, its salt, or its pharmaceutical composition may be administered by any route. In some embodiments, the novel compound, its salt, or its pharmaceutical composition is administered by various routes, including oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, subcutaneous, intracerebroventricular, transdermal, intradermal, rectal, intravaginal, intraperitoneal, topical (by powder, ointment, cream, and / or liquid drop), mucosal, nasal, oral, enteral, sublingual, intratracheal instillation, bronchial instillation, and / or inhalation, and / or oral spray, nasal spray, and / or aerosol. Particularly contemplated routes are systemic intravenous injection, topical administration by blood and / or lymphatic supply, and / or direct administration to the affected area. In general, the most suitable route of administration will depend on various factors, including the properties of the agent (e.g., its stability in the environment of the gastrointestinal tract) and the disorder of the subject (e.g., whether the subject can tolerate oral administration).

[0102] In the present invention, the amount administered to a child or adolescent may be determined by a specialist or a person skilled in the art, and may be less than or the same as that administered to an adult. The exact amount of the compound according to the present invention required to achieve an effective dose will vary from subject to subject, depending, for example, on the subject's species, age, and overall disorder, side effects or severity of the disorder, the identity of the specific compound, the mode of administration, etc.

[0103] It will be appreciated that the novel compounds and pharmaceutical compositions of the present invention can be used as combination therapy. The particular combination of therapies (therapeutic agents or procedures) to use in combination therapy will take into account the desired therapeutic effect to be achieved and the compatibility of the desired therapeutic agents and / or procedures.

[0104] The pharmaceutical composition of the present invention may be administered alone or in combination with one or more therapeutically active agents. In the case of "combination", the following delivery methods are within the scope of the present invention, but are not intended to suggest that the agents must be administered at the same time, administered or formulated for delivery at the same time. The composition may be administered simultaneously with, before, or after one or more other therapeutic agents or medical procedures of interest. Generally, each agent is administered at a dosage and / or time schedule established for that agent. Furthermore, the present invention includes delivery of the pharmaceutical composition of the present invention in combination with an agent that can improve its bioavailability, reduce and / or modify its metabolism, inhibit its secretion, and / or modify its distribution in the body. It will be better understood that the novel compound of the present invention and the therapeutically active agent used in this combination may be administered simultaneously in a single composition or separately in different compositions.

[0105] The particular combination to be used in the combination therapy will take into account the desired therapeutic effect to be achieved and / or the compatibility of the procedures and / or therapeutic active agents comprising the peptides of the invention. It will be understood that the combinations used can achieve the desired effect for the same disorder (e.g., the novel compounds of the invention may be administered in combination with yet another therapeutic active agent (e.g., a second therapeutic agent) used to treat the same disorder) and / or they can achieve a different effect (e.g., control of any side effects).

[0106] As used herein, the term "therapeutically active agent" refers to any substance used as a medicine to treat, prevent, delay, reduce or ameliorate a disorder, and refers to substances used in therapy, including prophylactic and curative treatments.

[0107] In some embodiments, the other therapeutically active agent for co-administration is for treating multiple myeloma. In some embodiments, the other therapeutically active agent may be a proteasome inhibitor and / or an immune modifying drug. In the present invention, the other therapeutically active agent may be selected from the group consisting of, but is not limited to, dexamethasone, bortezomib, carfilzomib, melphalan, doxorubicin, and cyclophosphamide.

[0108] In some embodiments, the pharmaceutical compositions of the present invention may be administered in combination with any therapeutically active agent or procedure (e.g., surgery, radiation therapy) useful for treating, alleviating, ameliorating, relieving, delaying the onset of, inhibiting the progression of, reducing the severity of, and / or reducing the incidence of one or more symptoms or characteristics.

[0109] In one aspect, the present invention can provide a kit for treating one or more diseases selected from the group consisting of inflammatory diseases and angiogenic diseases in a subject suffering from the disease.

[0110] In some embodiments, the kit includes (i) instructions for administering the novel compound or pharmaceutical composition according to the present invention to a subject suffering from the disease, and (ii) the novel compound or pharmaceutical composition according to the present invention. In some embodiments, the kit may include one or more unit dosage forms containing a dose of the novel compound or pharmaceutical composition described in the present invention effective to treat the disease in the subject. In some embodiments, the subject is a human patient.

[0111] In some embodiments, the kit further comprises at least one selected from the group comprising a sterile syringe, a sterile needle, a sterile IV bag, an infusion pump, or any combination thereof.

[0112] In another aspect, the present invention also provides a food composition containing a compound of formula 1:

[0113] [ka]

[0114] In the present invention, the food may be a health food with functional properties for alleviating or improving inflammation.

[0115] In the present invention, the food may be a health food for assisting in the treatment of angiogenic diseases.

[0116] In the present invention, the term "food composition" is used in a broad sense to encompass substances containing nutritional components, such as beverages, teas, energy drinks, alcoholic beverages, vitamin complexes, prebiotics, probiotics, postbiotics, dietary supplements, health functional foods and health foods, and is used to include not only all foods in the usual sense, but also "food additives" or "compositions for food additives" that are added to foods.

[0117] In the present invention, the food composition is characterized as being a functional health food having the function of alleviating or improving brain diseases.

[0118] The term functional food used in the present invention is synonymous with food for special health use (FoSHU), and refers to a food with high medical and therapeutic effects that is processed to efficiently exhibit a biological regulation function in addition to providing nutrition. Here, "functionality" means that it regulates nutrients to the structure and function of the human body or provides a useful effect for health purposes such as physiological action. The food of the present invention can be produced by a method commonly used in the art, and can be produced by adding raw materials and ingredients commonly added in the art. In addition, the dosage form of the food may be any dosage form recognized as a food, and the health functional food of the present invention may be in the form of powder, granules, tablets, capsules, or beverages.

[0119] The term "health food" refers to food that has a more active effect on maintaining or improving health than general foods, and "health supplement food" refers to food for the purpose of supplementing health. In some cases, the terms "health functional food," "health food," and "health supplement" are used interchangeably.

[0120] The composition may further contain a physiologically acceptable carrier, but the type of carrier is not particularly limited, and any carrier commonly used in the art can be used.

[0121] The composition may also contain additional ingredients commonly used in food compositions to improve aroma, taste, visual appearance, etc. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, etc. Minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr) may also be included. Furthermore, amino acids such as lysine, tryptophan, cysteine, and valine may also be included.

[0122] In addition, the composition may contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), bactericides (bleaching powder and highly bleached powder, sodium hypochlorite, etc.), antioxidants (butyl hydroxyanisole (BHA), butyl hydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color formers (sodium nitrite, sodium acetate, etc.), bleaching agents (sodium sulfite), seasonings (monosodium glutamate (MSG), etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavors (vanillin, lactones, etc.), leavening agents (alum, potassium D-hydrogen tartrate, etc.), strengthening agents, emulsifiers, thickeners (thickening agents), coating agents, gum bases, foam inhibitors, solvents, and improvers. The additives are selected according to the type of food and used in appropriate amounts.

[0123] The polysaccharide of the present invention may further contain a food-scientifically acceptable food supplement additive, or may be used with other foods or food ingredients, and may be used appropriately in a conventional manner. The amount of the active ingredient to be mixed may be appropriately determined depending on the purpose of use (prevention, health or therapeutic treatment).

[0124] The term "about" as used herein may be interpreted to mean approximately, roughly, roughly, or to a certain extent. When the term "about" is used in conjunction with a numerical range, it is interpreted to modify the range by extending the boundaries above and below the specified numerical values. In general, the term "about" is used herein to modify numerical values ​​above and below the stated value by a variance of 10%.

[0125] The terms "individual," "patient," and "subject" are used interchangeably and include any animal, including a mammal, e.g., mouse, rat, other rodents, rabbit, dog, cat, pig, cow, sheep, horse, or primate, including a human.

[0126] In the present invention, unless otherwise specified, "treatment" means reversing, alleviating, inhibiting the progression of, or preventing the disease or condition to which said term applies, or one or more symptoms of said disease or condition, and the term treatment as used herein refers to the act of treating, as "treating" is defined above. Thus, "treatment" or "therapeutic therapy" of a disease in a mammal may include one or more of the following: (1) Preventing the development of disease. (2) Prevent the spread of disease; (3) alleviate disease; (4) Preventing recurrence of disease; and (5) Palliating the symptoms of a disease In the present invention, unless otherwise specified, "prevention" refers to any action of suppressing or delaying the onset of an inflammatory disease or angiogenic disease by administering the pharmaceutical composition according to the present invention.

[0127] Pomalidomide used as a control in the present invention is (R,S)-4-amino-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione, and is characterized by being represented by [chemical formula 2].

[0128] [ka]

[0129] 3,6'-Dithiopomalidomide, which is used as a raw material in the synthesis of the present invention, can be represented by [Chemical Formula 3].

[0130] [ka]

[0131] Working Example The present invention will be described in more detail with reference to the following examples. It is obvious to those skilled in the art that these examples are merely for the purpose of illustrating the present invention and should not be construed as limiting the scope of the present invention.

[0132] Manufacturing Example A suspension of 3,6'-dithiopomalidomide (800 mg, 2.62 mmol, see US 9,623,020) in 52 ml DMSO and 24 ml H2O was heated at 100 °C for 17 h and HPLC (254 nm) analysis confirmed 3,6'-DP (0%), 3-MP (76%), and pomalidomide (24%). The deep red solution was cooled to room temperature and diluted with EtOAc. The solution was washed twice with H2O and once with brine. The aqueous layer was extracted once with EtOAc. The combined organic layers were dried over MgSO4, filtered, and concentrated. The residue was subjected to SiO2 column chromatography (Loading_silica gel that was dissolved in a mixture of acetone and a minimum amount of DMSO, coated with the residue, mixed with silica gel, and then rotary evaporated; Elution_CHCl3:acetone=20:1~9:1) to obtain 3-MP, which was further purified as a dark orange solid through a CHCl3 slurry followed by filtration (600 mg, 79% F005-02). The compound of [Formula 1] prepared in the present invention, i.e., 3-thiopomalidomide, can be abbreviated as 3-MP. [ka]

[0133] Example 1. In vitro CRBN binding assay Thalidomide-based compounds are known to regulate immune cell function and exert various pharmacological effects by binding to the E3 ligase cereblon (CRBN) and degrading the transcription factors Aiolos and Ikaros as its substrates. Based on this, the cereblon-binding ability of 3-MP was compared with that of pomalidomide in vitro.

[0134] The binding strength to cereblon was measured using the "AlphaScreen" method based on fluorescence resonance energy transfer (FRET), and the test was carried out using the PROTAC Optimization kit for BET Bromodomain-Cereblon binding kit (#79770, BPS bioscience, CA USA) according to the "Competitive Inhibition of the PROTAC assay" described in the kit manual. All reagents except for the candidate substance, DMSO, and Flag / Glutathione beads (#6765300, PerkinElmer, USA) were used as included in the kit.

[0135] As a result, as shown in Figure 1, it was confirmed that 3-MP binds to cereblon more effectively than pomalidomide, and the specific IC 50 The values ​​were 0.19 μM for 3-MP and 2.38 μM for pomalidomide, a difference of more than 10-fold.

[0136] Example 2. Evaluation of cytotoxicity The human multiple myeloma cell line MM.1S was treated with various concentrations of 3-MP and pomalidomide, and the cytotoxicity was evaluated.

[0137] MM.1S (CRL-2974) cell line, a human multiple myeloma cell line, was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured for 24 hours at 37°C in a 5% CO2 incubator using RPMI1640 medium (Corning, MD, USA) supplemented with 10% fetal bovine serum (Corning, MD, USA), 100 U / ml penicillin (Corning, MD, USA), and 100 μg / ml streptomycin (Corning, MD, USA). The cells were cultured at 1 × 10 4The cells were dispensed into a 96-well plate at 100 cells / well and cultured for 24 hours, after which they were treated with 0, 0.005, 0.014, 0.041, 0.123, 0.37, 1.111, 3.333, or 10 μM of pomalidomide or 3-MP, respectively, and cultured for an additional 3 days. After that, the cells were analyzed using a CCK analysis kit (Dojindo, Japan) according to the manufacturer's instructions.

[0138] As a result, as shown in Figure 2, the IC 50 IC of 0.069 μM, 3-MP 50 was confirmed to be 50 μM or higher, and 3-MP was found to have no cytotoxicity (blood toxicity) compared to pomalidomide.

[0139] Example 3. Mechanism of Cytotoxicity Reduction Thalidomide-related compounds are known to regulate immune cell function by binding to the E3 ligase cereblon (CRBN) and degrading the transcription factors Aiolos and Ikaros as substrates. We sought to determine whether 3-MP reduces the expression of Aiolos and Ikaros.

[0140] For this purpose, the MM.1S cell line was cultured in RPMI medium (Corning, MD, USA) supplemented with 10% fetal bovine serum (Corning, MD, USA), 100 U / ml penicillin (Corning, MD, USA), and 100 μg / ml streptomycin (Corning, MD, USA) at 37°C in a 5% CO2 incubator. The cells were cultured at 1 × 10 6After 24 h of incubation, cells were treated with 0.000128, 0.00064, 0.0032, 0.016, 0.08, 0.4, 2, or 10 μM pomalidomide and 3-MP, respectively, and incubated for 4 h. The cells were then disrupted with RIPA buffer containing a protease inhibitor cocktail (Thermo Fisher Scientific) and centrifuged at 14,000 rpm for 15 min at 4°C to obtain cell extracts. The cell extracts were loaded in equal amounts, separated using SDS-PAGE, and transferred to a PVDF membrane. The membrane with the transferred proteins was blocked with skim milk, incubated with primary antibodies for 3 h at room temperature, and then incubated with secondary antibodies with HRP for 1 h at room temperature. Between each step, the membrane was washed three times with TBS-T. Detection was performed by applying chemiluminescence reagents (Thermo Fisher Scientific) and confirmed using a Chemidoc (iBright CL1500, Invitrogen, CA, USA). Primary antibodies used were Aiolos (#15103), Ikaros (#9034), and GAPDH (#2118S) and secondary antibodies, which were purchased from Cell signaling technology (Danvers, MA, USA).

[0141] As a result, as shown in Figure 3a, it was confirmed that, unlike pomalidomide, 3-MP did not reduce the expression of Aiolos and Ikaros, which are substrates of cereblon.

[0142] Furthermore, the expression of Aiolos and Ikaros by 3-MP was confirmed at different times. 6After 24 h of incubation, cells were treated with 10 μM each of pomalidomide and 3-MP, and further cultured for 1, 4, 8, 12, or 24 h. The cells were disrupted with RIPA buffer containing a protease inhibitor cocktail (Thermo Fisher Scientific), and centrifuged at 14,000 rpm for 15 min at 4°C to obtain cell extracts. The cell extracts were loaded in equal amounts, separated using SDS-PAGE, and transferred to a PVDF membrane. The membrane with the transferred proteins was blocked with skim milk, incubated with primary antibody for 3 h at room temperature, and then incubated with secondary antibody with HRP for 1 h at room temperature. Between each step, the membrane was washed three times with TBS-T. Detection was performed using a chemiluminescence reagent (Thermo Fisher Scientific) and confirmed using a Chemidoc (iBright CL1500, Invitrogen, CA, USA). Aiolos (#15103), Ikaros (#9034), and GAPDH (#2118S) antibodies used as primary antibodies and secondary antibodies were purchased from Cell signaling technology (Danvers, MA, USA).

[0143] As a result, as shown in Figure 3b, it was confirmed that 3-MP did not reduce the expression of Aiolos and Ikaros, which are substrates of cereblon, more than pomalidomide.

[0144] Example 4. Mechanism of reducing reproductive toxicity Cereblon (CRBN)-binding therapeutics, such as thalidomide-based compounds, can induce severe congenital malformations, such as shortened forelimbs or brachydactyly, which have been reported to be closely related to the degradation of SALL4. Therefore, we compared the SALL4 degradation effect of 3-MP with that of pomalidomide.

[0145] For this purpose, human embryonic carcinoma cell line Tera-1 cell line (HTB-105) was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in a 37°C, 5% CO2 incubator using DMEM medium (Corning, MD, USA) supplemented with 10% fetal bovine serum (Corning, MD, USA), 100 U / ml penicillin (Corning, MD, USA), and 100 μg / ml streptomycin (Corning, MD, USA).

[0146] 5 x 10 cells 5 After 24 h of incubation, cells / well were dispensed into 12-well plates and treated with pomalidomide or 3-MP at 0.000128, 0.00064, 0.0032, 0.016, 0.08, 0.4, 2, or 10 μM, respectively, and incubated for an additional 4 h. Cells were disrupted with RIPA buffer containing a protease inhibitor cocktail (Thermo Fisher Scientific) and centrifuged at 14,000 rpm for 15 min at 4°C to obtain cell extracts. Equal amounts of cell extracts were loaded and separated using SDS-PAGE, and transferred to PVDF membranes. The membranes with transferred proteins were blocked with skim milk, incubated with primary antibodies for 3 h at room temperature, and then incubated with secondary antibodies with HRP for 1 h at room temperature. Between each step, the membranes were washed three times with TBS-T. Detection was performed by applying chemiluminescence reagents (Thermo Fisher Scientific) and confirmed using a Chemidoc (iBright CL1500, Invitrogen, CA, USA). SALL4 (#5850) and GAPDH (#2118S) antibodies used as primary antibodies and secondary antibodies were purchased from Cell signaling technology (Danvers, MA, USA).

[0147] As a result, as shown in Figure 4, 3-MP was found to cause less SALL4 degradation than pomalidomide, indicating that 3-MP is able to suppress teratogenic side effects more effectively than pomalidomide.

[0148] Example 5. Inflammation suppression effect Pomalidomide, an immunomodulatory agent, is known to enhance T cell-mediated immunity and natural killer (NK) cell-mediated immunity and suppress the production of pro-inflammatory cytokines (e.g., TNF-α, IL-1β, IL-6, and IL-8) by monocytes. Therefore, the inhibitory effect of 3-MP on the production of pro-inflammatory cytokines was compared with that of pomalidomide at the mRNA level.

[0149] For this, peripheral blood mononuclear cells (PBMCs, StemExpress, CA, USA) were plated in 96-well plates (Corning, MD, USA) at 1 × 10 6 The cells were dispensed at 100 cells / well and incubated in RPMI1640 (Corning, MD, USA) medium containing 10% fetal bovine serum (Corning, MD, USA) and 100 U / ml penicillin / streptomycin (Corning, MD, USA) at 37°C for one day for stabilization in a 5% CO2 incubator. Pomalidomide and 3-MP 10 mM DMSO stock (Sigma, USA) were diluted to 100 or 10 μM in the same medium as the PBMC culture solution and added to the corresponding PBMC wells (Final 10, 1 μM) and incubated at 37°C for one hour in a 5% CO2 incubator. All PBMC wells except for the non-inflammation group were added with 250 μg / μl (5% DMSO in RPMI1640) PHA (Sigma, USA) (Final 25 μg / μl) and incubated at 37°C for six hours in a 5% CO2 incubator.

[0150] PBMCs were prepped using an RNA prep kit (Monarch Total RNA Miniprep kit, NEB, UK) according to the kit manual, and the concentration was measured using a Nanophotometer (NP80, Implen, Germany). Depending on the concentration of each RNA sample, 1 μg of RNA was added, and cDNA was synthesized according to the manual of the cDNA synthesis kit (LunaScript RT SuperMix kit, NEB, UK).

[0151] The expression levels of each cytokine in the synthesized cDNA were measured by real-time PCR (StepOne TM The analysis was performed using a real-time PCR system (MicroAmp® Fast Optical 48-well plate, Applied Bioscience, USA). For the analysis, each well of a 48-well plate (MicroAmp® Fast Optical 48-well plate, Applied Biosystem, USA) contained 1 μl of cDNA, 10 pmol of forward and reverse primers for each gene, 8 μl of DW, and 10 μl of SYBR green (Luna® Master Mix, NEB, UK). The PCR reaction was performed according to the manual of the SYBR green kit, and the base sequences of the primers used are shown in Table 1. The gene-specific C of each sample obtained through real-time PCR were T Values ​​were normalized through the values ​​of the endogenous control gene (β-actin) for each sample, and 2ΔΔC T The expression level was back-estimated by calculation, and the result for each group was expressed as a relative quantification as a percentage value against the group treated with PHA alone.

[0152] [Table 1]

[0153] As a result, as shown in Figure 5a, 3-MP showed a similar level of pro-inflammatory cytokine suppression effect to that of pomalidomide.

[0154] We also confirmed the inhibitory effect of 3-MP on the production of proinflammatory cytokines at the mRNA level in mouse microglial cells (BV-2, ATCC, Manassas, VA, USA).

[0155] BV-2 was cultured in a 6-well plate (Corning, MD, USA) at 5 × 10 5The cells were dispensed at 100 cells / well and incubated in DMEM (Corning, MD, USA) medium containing 10% fetal bovine serum (Corning, MD, USA) and 100 U / ml penicillin / streptomycin (Corning, MD, USA) at 37°C in a 5% CO2 incubator for one day to stabilize. Pomalidomide and 3-MP 10 mM DMSO stock (Sigma, USA) were pretreated for one hour at concentrations of 30, 10, and 3.33 μM using the same medium as the BV-2 culture medium. All test groups, except for the non-inflammatory group, were added with 1 μg / mL LPS (Sigma, USA) and reacted for 6 hours at 37°C in a 5% CO2 incubator.

[0156] The drug-treated BV-2 was subjected to RNA prep using an RNA prep kit (Monarch Total RNA Miniprep kit, NEB, UK) according to the kit manual, and the concentration was measured using a Nanophotometer (NP80, Implen, Germany). Depending on the concentration of each RNA sample, 1 μg of RNA was added, and cDNA was synthesized according to the manual of the cDNA synthesis kit (LunaScript RT SuperMix kit, NEB, UK).

[0157] The expression levels of each cytokine in the synthesized cDNA were measured by real-time PCR (StepOne TM The analysis was performed using a PCR kit (Applied Bioscience, USA). For the analysis, each well of a 384-well plate (PCR Plate, 384-well, Thermo Fisher, USA) contained 1 μg of cDNA, 10 pmol of forward and reverse primers for each gene, 1 μl of DW, and 5 μl of SYBR green (Luna® Master Mix, NEB, UK). The PCR reaction was performed according to the manual of the SYBR green kit, and the base sequences of the primers used are shown in Table 2. The gene-specific C of each sample obtained through real-time PCR were T Values ​​were normalized through the values ​​of the endogenous control gene (GAPDH) for each sample, and 2ΔΔCT The expression levels were back-estimated by calculation, and the results for each group were expressed as a relative quantification using a percentage value relative to the group treated with LPS alone.

[0158] [Table 2]

[0159] As a result, as shown in Figure 5b, it was confirmed that 3-MP had a greater effect on suppressing pro-inflammatory cytokines than pomalidomide.

[0160] Example 6. Effect of suppressing oxidative stress To confirm the effect of suppressing oxidative stress, the expression levels of iNOS and COX-2 were examined.

[0161] For this purpose, the RAW264.7 cell line, a mouse macrophage cell line, was purchased from the Korea Cell Line Bank (KCLB, Korea) and used and cultured at 37°C in a 5% CO2 incubator using DMEM medium (Corning, MD, USA) supplemented with 10% fetal bovine serum (Corning, MD, USA), 100 U / ml penicillin (Corning, MD, USA), and 100 μg / ml streptomycin (Corning, MD, USA).

[0162] 2.4 x 10 RAW264.7 cells 5The cells were dispensed into 12-well plates at 100 cells / well, and after 24 hours of incubation, they were pretreated with pomalidomide and 3-MP at 3.33, 10, or 30 μM. After 1 hour, the cells were stimulated by treating with 60 ng / ml LPS, and then further cultured for 24 hours. The cells were disrupted with RIPA buffer containing a protease inhibitor cocktail (Thermo Fisher Scientific) and centrifuged at 14,000 rpm for 15 minutes at 4°C to obtain cell extracts. The cell extracts were loaded in equal amounts, separated by SDS-PAGE, and transferred to a PVDF membrane. The membrane with the transferred proteins was blocked with skim milk, incubated with primary antibody for 3 hours at room temperature, and then incubated with secondary antibody with HRP for 1 hour at room temperature. Between each step, the membrane was washed three times with TBS-T. Detection was performed by applying chemiluminescence reagents (Thermo Fisher Scientific) and confirmed using a Chemidoc (iBright CL1500, Invitrogen, CA, USA). The primary antibodies used, iNOS (#13120), COX-2 (#12282), and GAPDH (#2118), and the corresponding secondary antibodies, were purchased from Cell signaling technology (Danvers, MA, USA).

[0163] As a result, as shown in Figure 6a, it was found that 3-MP could suppress the expression of iNOS and COX-2 and effectively suppress oxidative stress, and its oxidative stress suppression effect was confirmed to be superior to that of pomalidomide.

[0164] To further confirm the inhibitory effect on oxidative stress, we examined the amount of nitrite released by mouse microglial cells (BV-2, ATCC, Manassas, VA, USA) in response to inflammatory responses, and the expression levels of iNOS, an enzyme that produces nitric oxide, and COX-2, an enzyme that produces cyclooxygenase-2.

[0165] To confirm the protein expression levels of nitrite, iNOS, and COX-2, BV-2 was plated in a 12-well plate (Corning, MD, USA) at 1.5 × 10 5The cells were dispensed at 100 cells / well and incubated in DMEM (Corning, MD, USA) medium containing 10% fetal bovine serum (Corning, MD, USA) and 100 U / ml penicillin / streptomycin (Corning, MD, USA) at 37°C for one day in a 5% CO2 incubator for stabilization. Pomalidomide and 3-MP 10 mM DMSO stock (Sigma, USA) were pretreated at 30, 10, and 3.33 μM for one hour in the same medium as the BV-2 culture medium. All test groups, except for the non-inflammatory group, were treated with 1 μg / mL LPS (Sigma, USA) and incubated for 24 hours in a 37°C, 5% CO2 incubator.

[0166] The cell culture medium was collected for nitrite measurement and centrifuged at 3,000 rpm for 5 min to obtain the supernatant. The nitrite concentration in the cell culture medium was measured using a microplate reader (NEO2MALPHAB Synergy Neo-2, BioTek, CA, US) according to the manual of the Griess Reagent System (#G2930, Promega, WI, USA).

[0167] Cells were harvested from the cell culture medium, disrupted with RIPA buffer containing a protease inhibitor cocktail (Thermo Fisher Scientific), and centrifuged at 14,000 rpm for 15 min at 4°C to obtain cell extracts. The cell extracts were loaded in equal amounts, separated using SDS-PAGE, and transferred to a PVDF membrane. The membrane with the transferred proteins was blocked with skim milk, incubated with primary antibodies for 3 h at room temperature, and then incubated with secondary antibodies with HRP for 1 h at room temperature. Between each step, the membrane was washed three times with TBS-T. Detection was performed by applying a chemiluminescence reagent (Thermo Fisher Scientific) and confirmed using a Chemidoc (iBright CL1500, Invitrogen, CA, USA). The primary antibodies used, iNOS (#13120), COX-2 (#12282), and GAPDH (#2118), and the corresponding secondary antibodies, were purchased from Cell signaling technology (Danvers, MA, USA).

[0168] As a result, 3-MP reduced the amount of nitrite released by LPS more effectively than pomalidomide (Figure 6b), and also reduced the expression of proteins related to oxidative stress (iNOS and COX-2) (Figure 6c), confirming that 3-MP is more effective at suppressing oxidative stress than pomalidomide.

[0169] Finally, to confirm the mRNA expression levels of iNOS and COX-2, BV-2 was cultured in a 6-well plate (Corning, MD, USA) at 5 × 10 5The cells were dispensed at 100 cells / well and incubated for one day in DMEM (Corning, MD, USA) containing 10% fetal bovine serum (Corning, MD, USA) and 100 U / ml penicillin / streptomycin (Corning, MD, USA) in a 5% CO2 incubator at 37°C for stabilization. Pomalidomide and 3-MP 10 mM DMSO stock (Sigma, USA) were pretreated for 1 hour at 30, 10, and 3.33 μM in the same medium as the BV-2 culture medium. All test groups, except for the non-inflammatory group, were treated with 1 μg / mL LPS (Sigma, USA) in a 37°C, 5% CO2 incubator for 6 hours.

[0170] The drug-treated BV-2 was subjected to RNA prep using an RNA prep kit (Monarch Total RNA Miniprep kit, NEB, UK) according to the kit manual, and the concentration was measured using a Nanophotometer (NP80, Implen, Germany). Depending on the concentration of each RNA sample, 1 μg of RNA was added, and cDNA was synthesized according to the manual of the cDNA synthesis kit (LunaScript RT SuperMix kit, NEB, UK).

[0171] The expression levels of each oxidative stress-related gene in the synthesized cDNA were measured using real-time PCR (StepOne TM The PCR reaction was carried out according to the manual of the SYBR green kit, and the base sequences of the primers used are shown in Table 3. The gene-specific Cs of each sample obtained through real-time PCR were analyzed and compared. Each well of a 384-well plate (PCR Plate, 384-well, Thermo Fisher, USA) was provided with 1 μg of cDNA, 1 μl of 10 pmol of forward and reverse primers for each gene, DW, and 5 μl of SYBR green (Luna® qPCR Master Mix, NEB, UK). The PCR reaction was carried out according to the manual of the SYBR green kit, and the base sequences of the primers used are shown in Table 3. T Values ​​were normalized through the values ​​of the endogenous control gene (GAPDH) for each sample, and 2ΔΔCT The expression levels were back-estimated by calculation, and the results for each group were expressed as a relative quantification using a percentage value relative to the group treated with LPS alone.

[0172] [Table 3]

[0173] As a result, as shown in Figure 6d, we confirmed that 3-MP could effectively suppress the mRNA expression of oxidative stress-related genes and had a superior oxidative stress inhibitory effect to pomalidomide.

[0174] Example 7.3 - Verification of MP in vivo stability Stock samples were prepared by dissolving the 3-MP standard in dimethyl sulfoxide (DMSO) at 1 mg / mL. To create a standard calibration curve, the stock sample was diluted in half with acetonitrile from 125 to 0.39 μg / mL to prepare standard solutions of different concentrations for all 10 samples, and the calibration curve was created as shown in Figure 7a.

[0175] In the case of test drugs for plasma stability verification, 3-MP was diluted to a concentration of 100 μg / mL and the experiment was carried out. For the plasma stability verification experiment, 10 μL of test drug manufactured at a concentration of 100 μg / mL was added to 90 μL human plasma, mixed for 30 seconds, and then incubated immediately after addition, for 15 minutes, 30 minutes, 60 minutes, and 120 minutes to prepare samples. Then, 400 μL of 100% ethyl acetate was added to each sample and extracted using a vortex mixer for 1 minute. The extracted samples were centrifuged at 13,000 rpm at 4 °C for 10 minutes, and the supernatant was collected and transferred to a new vial. The plasma extract sample was injected into a Hypersil GOLDTM C18 column (2.1 x 100 mm, 1.7 μm; Thermo Scientific, USA) coupled in-line with a UPLC-LTQ-Orbitrap 240 (Thermo Fisher Scientific, USA). The column temperature was 25°C, mobile phase A was a mixture of water and formic acid (100:0.1, v / v), and mobile phase B was a mixture of methanol and formic acid (100:0.1, v / v). Mobile phase B was initially increased from 5% to 95% for 9 minutes, and then maintained at 95% for 12 minutes. Mobile phase B was then lowered to 5% and equilibrated before being used for the next analysis. Analysis was performed with a mobile phase flow rate of 0.3 mL / min and a sample injection volume of 2 μL, and the mass spectrometer was operated in ESI positive mode. The spray voltage was set to 3.5 kV, and the flow rates of nitrogen sheath gas, auxiliary gas, and sweep gas were 50, 10, and 1 (arbitrary units), respectively. The capillary temperature was maintained at 250°C. Orbitrap data were collected in the range of m / z 100 to 1,000, and the analytical data were analyzed using Excalibur 4.0 software (Thermo Fisher Scientific, USA).

[0176] The 3-MP level in human plasma was confirmed by quantification based on the calibration curve in Figure 7a. As shown in Figure 7b, the 3-MP level in plasma was confirmed to be 80 μg / mL at 0 minutes, immediately after the addition of plasma, and although it showed a slight increase and decrease pattern over time, it was confirmed that 3-MP could be maintained in plasma without significant change from 0 minutes to 120 minutes.

[0177] As described above, certain parts of the present invention have been described in detail. It will be apparent to those skilled in the art that such specific techniques are merely preferred embodiments and do not limit the scope of the present invention. Therefore, the true scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. Compound represented by [Chemical Formula 1]: 【Chemistry 1】

2. A method for producing the compound of claim 1, comprising the steps of: (a) DMSO and H ２ 3,6'-dithiopomalidomide is dissolved in a 1,0-hexanediol mixed solvent and heated.

3. The mixed solvent in step (a) is DMSO:H ２ 3. The method according to claim 2, wherein O is mixed in a volume ratio of 1:0.1 to 1:

5.

4. 3. The method according to claim 2, wherein the heating in step (a) is performed at 80 to 120° C. for 5 to 30 hours.

5. The method according to claim 2, further comprising, after step (a), (b) purifying the compound according to claim 1.

6. The purification step (b) is carried out by diluting the solution produced in step (a) with EtOAc, drying the organic layer, and then removing the SiO ２ The method according to claim 5, further comprising purifying the product by column chromatography.

7. 10. A pharmaceutical composition for preventing or treating an inflammatory disease, comprising the compound according to claim 1 or a pharma- ceutically acceptable salt thereof as an active ingredient.

8. 8. The composition of claim 7, wherein the inflammatory disease is selected from the group consisting of psoriasis, rheumatoid arthritis and Crohn's disease.

9. 10. A pharmaceutical composition for preventing or treating angiogenic diseases, comprising the compound according to claim 1 or a pharma- ceutically acceptable salt thereof as an active ingredient.

10. The composition of claim 9, wherein the angiogenic disease is selected from the group consisting of corneal transplant angiogenesis, neovascular glaucoma, diabetic retinopathy, exudative age-related macular degeneration, diabetic macular edema, neovascular corneal disease, macular degeneration, pterygium, retinal degeneration, retrolental fibroplasia, granular conjunctivitis, hemangioma, angiofibroma, vascular malformation, arteriosclerosis, vascular adhesions and edema sclerosis.

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