KR20070005345A - 2,5-dihydro-4-alkyl-2,5-dioxofuran derivative and the use thereof - Google Patents

Abstract

2,5-Dihydro-4-alkyl-2,5-dioxofuran derivatives and use thereof are provided to improve inhibitory effects of T cell proliferation and IL-2(interleukin-2) secretion, so that the compounds are useful as immunosuppressant. The 2,5-dihydro-4-alkyl-2,5-dioxofuran derivatives represented by the formula(I), their derivatives or acid or base addition salts thereof are provided, wherein R1, R2, R3 and R4 are the same or different, and are each independently hydrogen, halogen atom, or a group selected from alkyl, hydroxy, alkoxy, polyhaloalkyl, carbonyl ester, cyanyl, nitro and amino. An immunosuppressant containing the composition for inhibiting T cell proliferation and IL-2 secretion comprises the 2,5-dihydro-4-alkyl-2,5-dioxofuran derivatives represented by the formula(I), their derivatives or acid or base addition salts thereof.

Description

2,5-dihydro-4-alkyl-2,5-dioxofuran derivatives and uses thereof {2,5-dihydro-4-alkyl-2,5-dioxofuran derivative and the use}

1 is a graph showing the IL-2 inhibitory effect of the compound of formula (I) of the present invention.

2 is a graph showing the MLR inhibitory effect of the compound of formula (I) of the present invention.

3 is a graph showing the IL-2 inhibitory effect of the compound of formula (II) of the present invention.

4 is a graph showing the MLR inhibitory effect of the compound of formula (II) of the present invention.

Field of technology

The present invention provides a 2,5-dihydro-4-alkyl-2,5-dioxofuran derivative of the formula (I) having T cell growth inhibition and IL-2 secretion efficacy, a method for preparing the same, and the same It relates to a pharmaceutical composition.

Prior art

The immune system is associated with signaling systems that recognize and eliminate pathogenic foreign substances such as viruses and bacteria. The living body distinguishes its cells or tissues (self-antigens) from foreign substances (non-self-antigens) and does not respond to self-antigens or exhibit immune function. On the other hand, the living body develops an acquired immune system that can efficiently remove nonmagnetic antigens introduced from foreign countries. In the immune system, T cells (T lymphocytes) are known to play an important role. In the peripheral, naive CD4 T cells are activated by receiving both antigen signals and other costimulatory molecule signals from antigen-presentation cells via T cell receptors. They then proliferate by secreting IL-2, a T cell proliferation factor. In this series of immune responses, when abnormal reactions occur, T cells can respond strongly to self-antigens and cause tissue damage. This is called autoimmune disease. Tissue damage or infection induced by autoimmune diseases results in various types of inflammatory reactions, tissue fibrosis or dysfunction. It is also therapeutically desirable to inhibit transplantation versus transplant disease due to organ transplant rejection or bone marrow transplantation, even if it is a normal immune response. These immune responses are carried out by the same mechanism, with activated T cells playing a major role. Thus, compounds having selective inhibitory activity against activated T cells have desirable efficacy as therapeutics for various diseases. Accordingly, in order to regulate mediators in T cells, compounds such as cyclosporin and FK-506 have been used. However, these compounds do not have sufficient selectivity for the activated T cells used so far, and side effects still remain.

The present inventors have tried to find a compound having a much better inhibitory effect than a compound having the effect of inhibiting T cell growth and inhibiting IL-2 (Interleukin-2) secretion, and is represented by the formula (I). The present invention was completed by confirming in animal models that hydro-4-alkyl-2,5-dioxofuran derivatives have very good cell growth inhibition and IL-2 secretion efficacy.

It is an object of the present invention to provide novel 2,5-dihydro-4-alkyl-2,5-dioxofuran derivatives having T cell growth inhibition and IL-2 secretion efficacy.

It is another object of the present invention to provide a process for the preparation of 2,5-dihydro-4-alkyl-2,5-dioxofuran derivatives.

Another object of the present invention is to provide a pharmaceutical composition comprising a 2,5-dihydro-4-alkyl-2,5-dioxofuran derivative compound and a pharmaceutically acceptable salt thereof.

The present invention also relates to immunosuppressive agents, in particular immunosuppressive agents which contain a selective inhibitor of activated T cells as an active ingredient. More specifically, the present invention relates to organ transplant rejection, bone marrow (hematopoietic stem) transplantation, graft-versus-graft disease, inflammatory response, autoimmune disease, or related diseases and fibrosis or dysfunction due to tissue damage or infection, Or to therapeutic agents for the treatment of diseases due to abnormal immune responses induced by activated T cells, such as allergic diseases.

Autoimmune diseases effectively prevented and treated by the immunosuppressive agents of the present invention include, for example, autoimmune hepatitis, rheumatoid arthritis, insulin dependent diabetes mellitus (IDDM), ulcerative colitis, multiple sclerosis (MS), scleroderma, severe Myasthenia gravis, multiple myositis / dermatitis, Hashimoto's disease, autoimmune cytopenia, Sjogren's syndrome, vasculitis syndrome or systemic lupus erythematosus.

In the present invention, allergic diseases include, for example, bronchial asthma, allergic rhinitis, atopic dermatitis, urticaria, hay fever and the like.

The present inventors synthesized the following formula (I) 2,5-dihydro-4-alkyl-2,5-dioxofuran derivative having T cell growth inhibitory activity and T cell IL-2 secretion inhibitory activity:

Formula (I)

Wherein R ₁ , R ₂ , R ₃ and R ₄ may be the same or different, each of which is independently a hydrogen atom, a halogen atom, or an alkyl, hydroxy, alkoxy, polyhaloalkyl, carbonyl ester, Group selected from cyanyl, nitro and amino.

The following compound of formula (II) is one derivative of formula (I):

Formula (II)

Wherein R ₁ , R ₂ , R ₃ and R ₄ may be the same or different, each of which is independently a hydrogen atom or an alkyl, hydroxy, alkoxy, polyhaloalkyl, carbonyl ester, halogen atom and Group selected from amino.

In the present invention, the term "alkyl" refers to a linear or branched hydrocarbon chain having 1 to 6 carbon atoms, and the term "alkoxy" refers to an alkyl-oxy group, wherein the alkyl chain is linear to 1 to 6 carbon atoms or Branched), and the term "alkylene" is a linear or branched divalent hydrocarbon chain of 1 to 6 carbon atoms, and the term "alkenylene" means 1 to 6 carbon atoms.

Pharmaceutically acceptable acids in the present invention include hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid, acetic acid, trifluoroacetic acid, lactic acid, pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, malic acid, Citric acid, ascorbic acid, methanesulfonic acid and camphoric acid, and the like, but is not limited thereto.

Pharmaceutically acceptable bases include, but are not limited to, sodium hydroxide, potassium hydroxide, triethylamine, and the like.

Compounds of formula (I) of the invention preferably are those wherein R ₁ is -OH; Compounds of formula (II) are preferably, R ₁ is an alkoxy or amino group; R ₂ is a hydrogen atom, a halogen atom or an alkyl, alkoxy, polyhaloalkyl or amino group, R ₃ is a hydrogen atom, a halogen atom or an alkyl, alkoxy, polyhaloalkyl or amino group, and R ₄ is a hydrogen atom, Alkyl, or polyhaloalkyl.

In the present invention, the formulas (Ia), (Ib), (Ic), (Id) and (Ie) are included in the compound of formula (I), and formulas (IIa) and (IIb) are included in formula (II).

The present invention also relates to a process for preparing the compounds of formulas (I) and (II) using the compound of formula (a) as starting material, which may or may not be purified according to conventional purification methods, It relates to a method for separating the stereoisomers in a conventional separation method and, if necessary, the conversion to addition salts with pharmaceutically acceptable acids or bases. In order to prepare the formula (I) of the present invention, the formula (Ia) (d in Scheme 1 below) is first synthesized using the compound of formula (a) as a starting material according to Scheme 1 below.

Scheme 1

Diethyl 2-{(2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) methyl} maronate of formula (Ia) synthesized as shown in Scheme 1 is shown in Scheme 2 According to formula (lb) (e in Scheme 2 below).

Scheme 2

In addition, diethyl 2-{(2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) methyl} maronate of formula (Ia) synthesized as shown in Scheme 1 The compounds of formula (IIa) (formula i in scheme 6) can be synthesized according to Schemes 3-6:

Scheme 3

Scheme 4

Scheme 5

Scheme 6

Further, 3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl)-of formula (Id) (formula g in scheme 4) synthesized as shown in Scheme 4 3-Hydroxy-propionic acid can be synthesized as a compound of formula (IIb) (formula j in scheme 7) according to Scheme 7:

Scheme 7

The present invention also relates to pharmaceutical compositions comprising, as active ingredient, one or more compounds of formula (I), derivatives thereof, alone or in combination with one or more pharmaceutically acceptable inert, non-toxic excipients or carriers. Pharmaceutical compositions of the present invention include, in particular, pharmaceutical compositions such as compositions suitable for parenteral or nasal administration, dragees or tablets, sublingual tablets, gelatin capsules, lozenges, suppositories, creams, ointments, and skin gels. May be mentioned. The effective dosage of the pharmaceutical composition of the present invention may vary depending on the weight and age of the patient, the nature and severity of the disease, and the route of administration in the case of oral, nasal, rectal or parenteral. Generally, a unit dose is administered at least once, from 0.1 to 250 mg per 24 hours.

Hereinafter, the present invention will be described according to Examples. The following examples are intended to illustrate the invention but do not limit the scope of the invention. The structure of the compounds described was confirmed by conventional spectroscopy and spectrometer techniques. Starting materials used in the present invention are known materials or products prepared according to known methods.

Example

Example 1-1: Diethyl 2-{(2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) methyl} maronate Synthesis

Step 1: Synthesis of 3,4-dimethylfuran-2,5-dione (1) [Formula b)]

Compound (a) was used as starting material. Maleanhydride (1 equiv) was dissolved in 98% acetic acid followed by the addition of 2-amino pyridine (2 equiv) and the decarbonation reaction was carried out. Until the CO ₂ gas evolution stops at reflux and heated to 150 ℃. In a state where no CO ₂ gas was generated, 2N sulfuric acid aqueous solution was added thereto, and the mixture was stirred at about 100 ° C. for about 1 hour. Distilled water was added to the reaction in the same volume, extracted with ethyl acetate, dried over anhydrous magnesium sulfate, and the mixed solvent was completely removed. After removing acetic acid and completely dried with a vacuum pump. The resulting solid was sufficiently dissolved in methylene chloride and then purified by silica acetate column using ethyl acetate: hexane (1: 3) flash column chromatography to give a clear solid as a product in about 67% yield.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 2.016 (s, 6H).

Step 2: Synthesis of 3- (bromomethyl) -4-methylfuran-2,5-dione Formula (c)

3,4-dimethylfuran-2,5-dione [Formula (b)] obtained in step 1 was used. Dimethylmaleanhydride (1 equiv), N-bromo succine imide (2 equiv) and benzoyl peroxide (0.167 equiv) were mixed and air was removed using argon from a reactor equipped with a reflux unit. Was completely removed and then heated to reflux at 76 to 78 ° C. for 5 hours while maintaining carbon tetrachloride at a concentration of 0.3 to 0.5 M. Then benzoylperoxide (0.167 equiv) was added and the reaction was held for 5 hours and then stirred at room temperature overnight. The reaction was filtered at room temperature, distilled water was added and extracted with ethyl acetate. Subsequent drying with anhydrous magnesium sulfate followed by complete removal of the mixed solvent. The resulting solid was dissolved in a small amount of methylene chloride and then purified by silica acetate column using ethyl acetate: hexane (1: 3) flash column chromatography to synthesize a transparent solid as a product in a yield of about 62%.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 4.157 (s, 2H), 2.156 (s, 3H)

Step 3: Synthesis of diethyl 2-{(2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) methyl} maronate [Formula (Ia)]

Form an anion with sodium hydrogen in tetrahydrofuran using dimethylmaronate, and then react with 3- (bromomethyl) -4-methylfuran-2,5-dione obtained in step 2 at -78 ° C. The product was obtained at a yield of at least%.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 4.203-4.085 (m, 4H) 3.811 (t, J = 7.8 Hz, 1H) 2.980 (d, J = 8.1 Hz, 2H) 2.085 (s, 3H) 1.250-1.191 (m, 6H)

Example 1-2: Synthesis of 3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) propionic acid [Formula (Ib)]

Diethyl 2-{(2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) methyl} maronate synthesized in Example 1-1 was 18%. It was mixed with dilute hydrochloric acid solution and heated to reflux for 12 hours. Then saturated aqueous sodium chloride solution was added, filtered and distilled water was added, followed by extraction with ethyl acetate. After drying over anhydrous magnesium sulfate, the mixed solvent was completely dried. The reaction was recrystallized from dichloromethane-petroleum ether (1: 2) to give the product in a yield of about 94%.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 2.748 (s, 4H) 2.101 (s, 3H)

Example 1-3 Synthesis of 3-bromo-3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) propionic acid [Formula (Ic)]

Br was introduced using a standard synthetic method using N-Bromosuccineimide (NBS) (E. Campaigne and B. F. Tuller, Organic Synthesis, 1963, Vol. IV, 921).

¹ H-NMR (300 MHz, CDCl ₃ ) δ 5.134-5.081 (m, 1H) 3.521 (d, J = 9.3 Hz, 1H) 3.387 (d, J = 6.9 Hz, 1H), 2.024 (s, 3H)

Example 1-4: Synthesis of 3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) -3-hydroxy-propionic acid [Formula (Id)]

The bromo compound f was cooled in an ice water vessel and then added to the 1N KOH solution and stirred for 1 hour. The pH was adjusted to 1 with 3N HCl, and extracted with ethyl acetate. The mixed solvent was dried completely using anhydrous magnesium sulfate and then additional solvent was added. The residue was separated by chromatography using a methanol: dichloromethane (1: 5) solution to give the title compound in 15% yield as product.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 4.146-4.075 (m, 1H) 2.752 (s, 2H) 2.089 (s, 3H)

Example 1-5: Synthesis of methyl 3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) propionate [Formula (Ie)]

The compound of Examples 1-4 (20 mg, 0.11 mmol) was dissolved in a benzene: dichloromethane (3: 1) solution. TMS- diazomethane (trimethylsilyldiazomethane) (14 mg, 1.2 equivalents) was added, and after about 10 minutes of discoloration, a small amount of water was added, followed by extraction with ethyl acetate. The organic layer was dried over magnesium sulfide and the solution was removed. The residue was separated by chromatography using an ethylacetate: hexanes (1: 3) solution to give the title compound in product 92% yield (20 mg) as a product.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 4.34 (s, 3H), 2.72 (t, J = 6.9 Hz, 2H), 2.54 (t, J = 7.5 Hz, 2H), 1.98 (s, 3H)

Example 1-6: Synthesis of (Z) -trimethyl-pent-3-ene-1,3,4-tricarboxylate [Formula (IIa)]

The compound of Example 1-4 (25 mg, 0.136 mmol) was dissolved well in a methanol: dichloromethane (1: 1) solution and then EDC {1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride} (31.24 mg, 1.3 equiv) and dimethylaminopyridine (5 mg, 0.3 equiv) were reacted at 0 ° C. and argon gas for about 1 hour. After the reaction was completed by adding a small amount of water, the mixture was extracted with ethyl acetate, and the organic layer was dried over magnesium sulfide and the solution was removed. The residue was separated by chromatography using an ethylacetate: hexanes (1: 3) solution to give the title compound as a product in 39% yield (13 mg).

¹ H-NMR (300 MHz, CDCl ₃ ) 4.59 (s, 3H), 4.57 (s, 3H), 4.55 (s, 3H), 2.73 (t, J = 6.9 Hz, 2H), 2.55 (t, J = 7.5 Hz, 2H), 1.98 (s, 3H)

Example 1-7: Synthesis of (Z) -trimethyl-pent-3-ene-1,3,4-tricarboxylate [Formula (IIb)]

The compound of Examples 1-4 (27 mg, 0.146 mmol) was dissolved well in a methanol: dichloromethane (1: 1) solution and then EDC {1-ethyl-3- (3-dimethylaminopropyl) -carbodiimide hydrochloride} (31.24 mg, 1.3 equiv) and dimethylaminopyridine (5 mg, 0.3 equiv) were reacted at 0 ° C. and argon gas for about 1 hour. After adding a small amount of water to terminate the reaction, the mixture was extracted with ethyl acetate and the organic layer was dried over magnesium sulfide and the solution was removed. The residue was separated by chromatography using an ethylacetate: hexane (1: 3) solution to give the title compound as a product in 20% yield (20 mg).

¹ H-NMR (300 MHz, CDCl ₃ ) 7.35-7.26 (m, 15H), 5.09 (s, 2H), 5.06 (s, 2H), 5.04 (s, 2H), 2.71 (t, J = 6.9 Hz, 2H), 2.52 (t, J = 7.5 Hz, 2H), 1.97 (s, 3H)

Example 2: Pharmacological Activity

Example  2-1: IL-2 Secretion Inhibitory Effect of the Compound of Formula (I)

For the compound of formula (I) synthesized in Example 1-2, ELISA was performed using a culture solution containing the compound to investigate the efficacy of IL-2 secretion inhibition in Jurket cells. Human T cancer cells (from Yonsei University) were suspended in RPMI 1640 medium (gibco) containing 10% fetal bovine serum, 2 mM L-glutamine and 100 unit penicillin, followed by 2 x Cells were plated to 10 ⁵ (cells / well) and cells were activated using 1 μM ionomycin and 50 ng PMA (phorbol 12-myristate 13-acetate). The compounds of formula (I) of the following Examples 1-2 were treated at concentrations of 10, 1, ^10-1 and ^10-2 μg / ml respectively and incubated for 24 hours. After incubation, 100 μl of the supernatant obtained by centrifugation was respectively dispensed into wells of a plate coated with antibody, and reacted for 2 hours, followed by 100 μl of working detector (Detection antibody + avidin-HRP reagent). Was added and reacted for 1 hour. After addition of the substrate solution (Tetramethylbenzidine, TMB), the reaction was terminated with 2N H ₂ SO ₄ solution and the values of OD 570-450 nm were recorded. As a result, as shown in FIG. 1, the IC ₅₀ for IL-2 of the compound of formula (I) was 5.29 μM, demonstrating that it effectively inhibits IL-2 secretion of T cells.

Example 2-2 Determination of Mixed Lymphocyte Reaction (MLR) of a Compound of Formula (I)

The compound of Example 1-2 was used. The drug effect on MLR was measured to measure the immune response of intercellular heterologous antigens. Lymphocytes were isolated from the spleen of Balb / c and C57BL / 6 mice (obtained from central laboratory animals). Responsive cells were prepared by floating 5 x 10 ⁶ / ml in RPMI 1640 medium (hereinafter referred to as "complete medium") to which splenocytes of C57BL / 6 mice were added with 10% fetal calf serum. Splenocytes of BALB / c mice used as stimulator cells were washed twice with HBSS (Hanks' balanced salt solution) after 50 ug / ml mitomycin C treatment, and then suspended in complete medium at 1 x 10 ⁷ / ml. . The negative control group was prepared by adding 50 μl of reaction cells and stimulatory cells in two independent wells, and then adding 150 μl of complete medium to each well. The experimental control group was treated with 50 μl of reaction cells and stimulation cells in one well, and then 100 μl of complete medium was used for the experiment. The experimental group serially dilutes the compound of formula (I) in PBS (10, 1, ^10-1 , ^10-2 , ^10-3 and ^10-4 μg / ml), and adds 2 μl of this solution to each well. 100 μl of complete medium was added so that the total amount was 200 μl per well. All plates were incubated for 72 hours at 37 ° C., 5%, CO ₂ thermohygrostat (hereinafter referred to as “incubator”), and 10 μl (0.1 Ci) of 3H-TdR (Tritiated thymidine, 20.0 Ci / mmol), respectively. It was added to the wells and incubated for 6 hours. After incubation, the cells of each well were adsorbed onto a glass filter fiber with a cell harvester and dried, and then the amount of ³ H-TdR incorporated into the cell was measured using a scintillation beta counter. By measuring the proliferation inhibitory response of T cells. As a result, as shown in FIG. 2, the compound of formula (I) of the present invention showed almost the same level of T cell growth inhibitory effect as cyclosporin A, which is widely used as an immunosuppressant.

Example 2-3 Inhibitory Effect on Organ Transplant Rejection of Compound of Formula (I)

The compound of Example 1-2 was used for the experiment. The inhibitory effect of organ transplant rejection on compound (I) was investigated. Anesthetized male Lewis rats (obtained from central animals) from 8 to 10 weeks of age as cardiac donor rats, followed by an incision in the abdomen and rib cage, and 50 units (2 ml) of heparin through the IVC. After perfusion, the lower process vein and the upper process vein (SVC) were bled sequentially, and 4 ° C saline was injected into the aorta and perfused to the myocardium through the coronary artery. Next, while observing the color of the heart, when the color of the heart turned white, the aorta, the pulmonary artery, the upper process vein and both lungs were cut in turn to extract the heart, and stored in 4 ℃ physiological saline. Next, 10-week-old male Wistar rats (obtained from central animals) were anesthetized as cardiac rats, and the abdominal cavity was dissected to expose the abdominal aorta and the lower step vein, followed by a Satinsky clamp. Block the aorta and the aorta together, and using a 10-0 polypropylene suture under a surgical microscope, the aorta of the donor's heart and the abdominal aorta of the donor -Side sutures. Subsequently, regular contractions of the atria and ventricles were observed through ventricular fibrillation by reperfusion, and the abdominal wall was sutured with a suture. After dissolving the compound of formula (I) in DMSO at a concentration of 10 mg / ml, PBS (phosphate-buffered saline, 8.0 g NaCl, 1.16 g Na ₂ HPO ₄ , 0.2 g KH ₂ PO ₄ , 0.2 g KCl, 1 L water) Injectable preparations were prepared by diluting to concentrations of 0.5 mg / ml and 1.0 mg / ml, and after 15 intravenous injections for 14 days from the day of transplantation, the heartbeat was palpated. When the heartbeat was not palpated for at least two times, it was determined that the transplantation heart had a rejection phenomenon. As a result, as shown in Table 1 below, survival of 150 days or more was shown at a dose of 0.1 to 0.5 mg / kg of the compound of formula (Ib) of Example 1-2. This result demonstrates that the compound of formula (I) of the present invention has very good organ rejection reaction inhibitory effect compared to that of cyclosporin A at the dose of 5 mg / kg.

Table 1.

Substance Dosage Number of transplants (horses) Average survival Saline solution - 10 7.6 ± 1.6 Formula (Ib) Compound 0.1 mg / kg 10 176.4 ± 11.3 0.5 mg / kg 10 183.7 ± 9.4

Example 2-4: Inhibitory Effect of the Compound of Formula (II) on IL-2 Secretion

The inhibitory effect of IL-2 secretion was analyzed for the compound of formula (II) of Example 1-7. As a result, as shown in FIG. 3, the IC ₅₀ values of the compound of formula (II) and cyclosporin A were 4.22 μM and 9.40 μM, respectively. This indicates that the compound of formula (II) has a very good inhibitory effect of IL-2 secretion.

Example 2-5 MLR Inhibitory Effects of Compounds of Formula (II)

In the same manner as in Example 2-2, the effect of MLR inhibition on the compound of formula (II) was analyzed. As shown in FIG. 4, the results of the IL-2 and MLR inhibitory effects of the compound of formula (II) derived from formula (I) demonstrate that the compound of formula (II) has a very good effect as an immunosuppressive agent. .

Example 2-6 Acute Toxicity Test of Compounds of Formula (I)

The compound of Example 1-2 was used. The compound of formula (Ib) was dissolved in DMSO and injected into three treatment groups of 10, 50 and 100 mg / kg concentrations in the veins of 5 male and female ICR mice (Samtaco), respectively, and observed changes. It was. As a result, two rats died in the high-dose 100 mg / kg injection group. Thus, the anti-lethal amount (LD ₅₀ ) of the compound of formula (I) was determined to be 100 mg / kg or more. The half dose of 100 mg / kg is 1,000 times the effective dose of 0.1 mg / kg.

The 2,5-dihydro-4-alkyl-2,5-dioxofuran derivative of the present invention is a compound having very good T cell growth inhibition and IL-2 (Interleukin-2) secretion efficacy, Can be used for development.

Claims (6)

Formula (I) compounds, derivatives thereof or acid or base addition salts thereof: Formula (I)

Wherein R ₁ , R ₂ , R ₃ and R ₄ may be the same or different, each of which is independently a hydrogen atom, a halogen atom, or an alkyl, hydroxy, alkoxy, polyhaloalkyl, carbonyl ester, Group selected from cyanyl, nitro and amino. A compound according to claim 1, wherein R ₁ is -OH. The method of claim 1, Diethyl 2-{(2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) methyl} maronate; 3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) propionic acid; 3-bromo-3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) propionic acid; 3- (2,5-Dihydro-4-methyl-2,5-dioxofuran-3-yl) -3-hydroxy-propionic acid; Methyl 3- (2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) propionate; (Z) -trimethyl-pent-3-ene-1,3,4-tricarboxylate; And And (Z) -trimethyl-pent-3-ene-1,3,4-tricarboxylate. Formula I, characterized in that it comprises a diethyl 2-{(2,5-dihydro-4-methyl-2,5-dioxofuran-3-yl) methyl} maronate synthesis process according to Scheme 1 Method for preparing the compound of Scheme 1

A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable acid or base addition salt thereof, as an active ingredient, alone or in admixture with one or more pharmaceutically acceptable excipients. A pharmaceutical composition having cell growth inhibition and IL-2 secretion inhibitory activity. The pharmaceutical composition according to claim 5, which is an immunosuppressive agent.

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