KR20060031956A - Pharmaceutical composition for the prevention and treatment of liver fibrosis and cirrhosis through direct kinase activity of ｐ90 ribosomal S6 kinase 1 (RSS1) - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating hepatic fibrosis and cirrhosis by directly enhancing the kinase activity of p90 ribosomal S6 kinase 1 (RSK1) by small molecule organic compounds, wherein the direct increase of kinase activity of RSK1 by small molecule organic compounds is CCAAT-. Enhancement of hepatic fibrosis and cirrhosis can be improved or inhibited through the inhibition of expression of transforming growth factor (β) through selective phosphorylation of the serine residue of the enhancer binding protein β protein. Furthermore, organic compounds that enhance the activity of RSK1 promote the regeneration of hepatocytes and thus can be used as therapeutic agents for liver fibrosis and cirrhosis.

p90 ribosomal S6 kinase, RSK1, treating cirrhosis

Description

The organic agents that directly activate p90 ribosomal S6 kinase 1 (RSK1) for the use of prevention and treatment of liver fibrosis by directly promoting kinase activation of ribosomal S6 kinase 1 (RSS1). and cirrhosis}

Figure 1 shows that the activity of RSK1 in H4IIE cells by the typical small-molecule organic compound Oltipraz,

2 shows that RSK1 activity is directly enhanced by oltipraz, which is a representative small molecule organic compound.

3 is a diagram showing an example of a possible model in which oltipraz, a representative dithiol-based small molecule organic compound, can be combined with RSK1,

FIG. 4 shows a possible model of another small molecule organic compound, a thiophene derivative, which may bind to RSK1, and similarly to oltipraz, a thiophene derivative directly activates RSK1.

FIG. 5 shows that direct activation of RSK1 by a typical small-molecule organic compound, Oltipraz, results in selective phosphorylation of the 105-serine residue of CCAAT-enhancer binding factor β (C / EBPβ) protein, a cellular regeneration factor. CTT-RSK1, K112 / 464R-RSK1 (CTT-RSK1, K112 / 464R-RSK1), which show that nuclear translocation and DNA binding capacity of EBPβ are increased, induction of C / EBPβ is increased, and that phosphorylation of C / EBPβ, nuclear transport and gene expression of C / EBPβ are caused by RSK1 activity. RSK1 variant) is shown to be suppressed by overexpression,

FIG. 6 shows that representative small molecule organic compounds directly increasing the activity of RSK1 inhibit C / EBPβ dependent gene expression of transforming growth factor β (TGFβ), which is a key mediator of liver fibrosis and cirrhosis. ,

7A to 7B show the results of searching for GST-induced expression by C / EBPβ activation and C / EBPβ activation by a novel synthesized compound having an oltipraz-like action,

8A to 7C are tables listing chemical names of newly synthesized compounds,

FIG. 9 is a diagram showing a pathway for treating hepatic fibrosis and cirrhosis through the inhibition of RSK1 activation by small molecule organic compounds through the inhibition of expression of conversion growth factor through specific site phosphorylation of C / EBβ.

The present invention relates to a composition of small-molecule organic compounds including oltipraz, which can prevent and treat liver fibrosis and cirrhosis through direct activation of RSK1. In particular, in the present invention, p90 ribosomal S6 kinase 1 (RSK1) may be directly activated by small molecule organic compounds to enhance kinase activity, thereby forming active C / EBPβ by phosphorylating characteristic residues of C / EBPβ, As a result, gene expression is regulated to promote hepatocyte regeneration, and active C / EBPβ inhibits the gene expression of transforming growth factor β (TGFβ), a key factor in liver fibrosis, thereby preventing liver fibrosis and cirrhosis. Can be prevented and treated.

The liver plays a pivotal role in metabolism in the body and in the body, and is a living organ that continuously undergoes enzyme reactions and energy metabolism. Currently, hepatitis, cirrhosis and liver cancer account for the highest proportion of chronic diseases in Korea, along with circulatory disease, and account for a large proportion of the causes of death due to diseases. In particular, the drinking population is relatively high compared to the developed countries, and the cause of liver damage caused by binge drinking is high. Continuous liver tissue damage caused by viral infections or drinking alcohol is characterized by chronic complications that lead to cirrhosis or liver cancer. Considering the physiological characteristics and importance of liver tissue, the treatment and prevention of liver disease is very important, and it is necessary to identify and utilize the therapeutic action points of drugs that can reduce liver tissue damage and ultimately normalize liver function through liver cell regeneration. Invention is required.

The present inventors have revealed through the previous studies that the activity of C / EBP is an essential core transcription factor in the treatment of cirrhosis and liver tissue regeneration (Korean Patent Application No. 2000-18134 and Korean Patent No. 0377789). Accordingly, the present inventors have repeatedly conducted research on a technique for searching for cellular signals mediating changes in major gene expressions related to liver fibrosis and cirrhosis, and as a result, small-molecule organic compounds can directly enhance the kinase activity of RSK1. And the direct increase of RSK1 activity to treat liver fibrosis, cirrhosis of the liver, and to complete a pharmaceutical composition containing a small molecule organic compound that can directly enhance the kinase activity of RSK1.

Accordingly, an object of the present invention relates to a composition of small molecule organic compounds for treating and preventing hepatic fibrosis, cirrhosis through enhancement of kinase activity of RSK1.

In the present invention, p90 ribosomal S6 kinase 1 (RSK1) may be directly activated by small molecule organic compounds to enhance kinase activity, thereby forming active type C / EBPβ by phosphorylating characteristic residues of C / EBPβ. As a result, gene expression is regulated, which not only promotes the regeneration of hepatocytes, but also enhances the kinase activity of RSK1 directly by small-molecule organic compounds through selective phosphorylation of the 105 serine residue of CCAAT-enhancer binding protein β protein. The present invention relates to a pharmaceutical composition for preventing and treating liver fibrosis and cirrhosis, which can directly activate RSK1 because it can improve or treat liver fibrosis and cirrhosis through suppression of expression of factor-β).

In order to achieve the above object, small molecule organic compounds which directly increase the kinase activity of RSK1 include organic compounds including 1,2-dithiole-3-thione and a bicyclic ring molecule structure and derivative organic compounds thereof. Bicyclic ring molecules may contain pyrazine, pyridazine, pyrimidine, thiazole, thiopene (Table 1).

In the present invention, diseases that can be prevented and treated by the present technology include hepatic fibrosis and cirrhosis diseases. In the present invention, the small molecule organic compound includes derivative compounds having various structures such as pyrazine, pyridazine, pyrimidine, thiazole, and chiphene, including 1,2-dithiole-3-thione and a bicyclic ring molecule structure. Bicyclic ring molecular organic compounds include, but are not limited to the following compounds.

Table 1. Examples of representative 1,2-dithiole-3-thione and bicyclic ring molecules containing RSK1 activator

In the formula (7),

X means carbon or nitrogen,

Q means sulfur, oxygen or -S = O,

R ¹ and R ² are each independently hydrogen, C _1-7 -alkyl, C _3-7 -cycloalkyl, C _1-7 -haloalkyl, C _1-7 -alkoxy, C _3-7 -cycloalkoxy, C _1-7 -alkylthio, C _3-7 - cycloalkyl alkylthio, C _1-7 - alkenyl, C _1-7 alkynyl, C _1-7 -alkyl sulfonyl, C _1-7 -alkyl-aminocarbonyl , HO-C _1-7 -alkyl, HS-C _1-7 -alkyl, hydroxy, thiol, halogen, carboxyl, nitro, cyano, C _1-7 -alkylcarbonyl, C _1-7 -alkoxycar Carbonyl, C _1-7 -alkylcarbonyloxy, C _{1-7 -alkylcarbonyl} -C _1-4 -alkyl, C _1-4 -alkoxy-C _1-4 -alkyl, C _1-4 -alkylthio- C _1-4 -alkyl, amino, C _1-7 -alkylamino, C _1-7 -alkylcarbonylamino, C _1-4 -alkoxy-C _1-4 -alkylamino, C _1-4 -alkylthio- C _1-4 -alkylamino, C _1-4 -alkylsulfonamino, phenyl, heteroaryl, phenyl-C _1-4 -alkyl, heteroaryl-C _1-4 -alkyl, phenyl-C _1-4 -alkoxy- C _1-4 -alkyl, phenyl-C _1-4 -alkylthio-C _1-4 -alkyl, phenoxy-C _1-4 -alkyl, phenylthio-C _1-4 -alkyl, Phenylcarbonylamino, phenoxy-C _1-4 -alkylcarbonylamino, phenyl-C _1-4 -alkoxy-C _1-4 -alkylcarbonylamino, heteroaryloxy-C _1-4 -alkyl, heteroaryl Thio-C _1-4 -alkyl, heteroaryl-C _1-4 -alkylthio-C _1-4 -alkyl.

On the other hand, the heteroaryl means a 5 or 6 membered ring containing one or more heteroatoms selected from the group consisting of nitrogen, sulfur and oxygen.

Further, the phenyl and heteroaryl may be substituted or unsubstituted, and possible substituents include halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _1-7- Alkylthio, C _1-7 -alkenyloxy, C _1-4 -alkylcarbonyl, C _1-4 -alkylamino, nitro, amino, cyano, HO-C _1-4 -alkyl, HS-C _{1- 4} -alkyl, HO-C _1-7 -alkoxy, HO-C _1-7 -alkylthio, HS-C _1-7 -alkylthio, HS-C _1-7 -alkoxy, consisting of thiols, hydroxy, carboxyl groups Selected from the group, these may be mono- or polysubstituted.

Meanwhile, the phenyl and heteroaryl may be each independently fused with one or more benzene or heteroaryl having the same meaning.

In addition, the fused phenyl and heteroaryl may be substituted or unsubstituted, and possible substituents include halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _{1- 7} -alkylthio, C _1-7 -alkenyloxy, C _1-4 -alkylcarbonyl, C _1-4 -alkylamino, nitro, amino, cyano, HO-C _1-4 -alkyl, HS-C _1-4 -alkyl, HO-C _1-7 -alkoxy, HO-C _1-7 -alkylthio, HS-C _1-7 -alkylthio, HS-C _1-7 -alkoxy, thiol, hydroxy, carboxyl group It is selected from the group consisting of, these may be mono- or polysubstituted.

In the formula (8),

Q means sulfur or oxygen,

Y, Z, W and L each independently represent carbon, nitrogen, oxygen or sulfur,

R ³ , R ⁴ , R ⁵ , R ⁶ are each independently hydrogen, C _1-7 -alkyl, C _3-7 -cycloalkyl, C _1-7 -haloalkyl, C _1-7 -alkoxy, C _{3- 7} -cycloalkoxy, C _1-7 -alkylthio, C _3-7 -cycloalkylthio, C _1-7 -alkyl sulfonyl, C _1-7 -alkylaminocarbonyl, hydroxy, thiol, halogen, carboxyl , Nitro, cyano, C _1-7 -alkylcarbonyl, C _1-7 -alkoxycarbonyl, C _1-7 -alkylcarbonyloxy, amino, C _1-7 -alkylamino, C _1-7 -alkyl Carbonylamino, C _1-4 -alkylsulfonamino, phenyl, heteroaryl, phenyl-C _1-4 -alkyl, heteroaryl-C _1-4 -alkyl.

On the other hand, the heteroaryl means a 5 or 6 membered ring containing one or more heteroatoms selected from the group consisting of nitrogen, sulfur and oxygen.

Further, the phenyl and heteroaryl may be substituted or unsubstituted, and possible substituents include halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _1-7- Alkylthio, C _1-4 -alkylcarbonyl, C _1-4 -alkylamino, nitro, amino, cyano, thiol, hydroxy, carboxyl groups, which may be mono- or polysubstituted.

Meanwhile, the phenyl and heteroaryl may be each independently fused with one or more benzene or heteroaryl having the same meaning.

In addition, the fused phenyl and heteroaryl may be substituted or unsubstituted, and possible substituents include halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _{1- 7} -alkylthio, C _1-4 -alkylcarbonyl, C _1-4 -alkylamino, nitro, amino, cyano, thiol, hydroxy, carboxyl groups, which may be mono- or polysubstituted have.

Meanwhile, the six-membered ring including Y, Z, W, and L of Formula 2 may be fused with one or more phenyl and heteroaryl.

In the formula (9),

Q means sulfur or oxygen,

A, D, and E each independently represent carbon, nitrogen, oxygen, or sulfur,

R ⁷ , R ⁸ , R ⁹ are each independently hydrogen, C _1-7 -alkyl, C _3-7 -cycloalkyl, C _1-7 -haloalkyl, C _1-7 -alkoxy, C _3-7 -cyclo Alkoxy, C _1-7 -alkylthio, C _3-7 -cycloalkylthio, C _1-7 -alkylsulfonyl, C _1-7 -alkylaminocarbonyl, hydroxy, thiol, halogen, carboxyl, nitro, Cyano, C _1-7 -alkylcarbonyl, C _1-7 -alkoxycarbonyl, C _1-7 -alkylcarbonyloxy, amino, C _1-7 -alkylamino, C _1-7 -alkylcarbonylamino , C _1-4 -alkylsulfonamino, phenyl, heteroaryl, phenyl-C _1-4 -alkyl, heteroaryl-C _1-4 -alkyl.

On the other hand, the heteroaryl means a 5 or 6 membered ring containing one or more heteroatoms selected from the group consisting of nitrogen, sulfur and oxygen.

Further, the phenyl and heteroaryl may be substituted or unsubstituted, and possible substituents include halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _1-7- Alkylthio, C _1-4 -alkylcarbonyl, C _1-4 -alkylamino, nitro, amino, cyano, thiol, hydroxy, carboxyl groups, which may be mono- or polysubstituted.

Meanwhile, the phenyl and heteroaryl may be each independently fused with one or more benzene or heteroaryl having the same meaning.

In addition, the fused phenyl and heteroaryl may be substituted or unsubstituted, and possible substituents include halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _{1- 7} -alkylthio, C _1-4 -alkylcarbonyl, C _1-4 -alkylamino, nitro, amino, cyano, thiol, hydroxy, carboxyl groups, which may be mono- or polysubstituted have.

Meanwhile, the 5-membered ring including A, D, and E of Formula 3 may be fused with one or more phenyl and heteroaryl.

In the formula (10),

Q means sulfur or oxygen,

R ¹⁰ is hydrogen, halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _1-7 -alkylthio, C _1-4 -alkylcarbonyl, C _1-4 -Alkylamino, nitro, amino, cyano, thiol, hydroxy, carboxyl groups, which may be mono- or polysubstituted as substituents of phenyl.

In the formula (11),

n = 0, 1, or 2,

R ¹¹ is C _1-4 -alkyl,

R ¹² and R ¹³ is C _1-4 each independently is selected from the group consisting of alkyl, - alkyl, C _1-4

R ¹⁴ is hydrogen, halogen, C _1-7 -alkyl, C _1-7 -alkoxy, C _1-7 -haloalkyl, C _1-7 -alkylthio, C _1-4 -alkylcarbonyl, C _1-4 -Alkylamino, nitro, amino, cyano, thiol, hydroxy, carboxyl groups, which may be mono- or polysubstituted as substituents on the pyrazine nucleus.

Unless stated otherwise, the above definitions are used as follows.

Halo is the generic name for fluoro, chloro, bromo and iodine;

C _1-4 -alkyl is a straight and branched chain saturated hydrocarbon radical having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, n-butyl, 1-methylethyl, 2-methylpropyl, 1,1-dimethylethyl And the like;

C _1-7 -alkyl comprises higher order homologues comprising C _1-4 alkyl (as defined above) and containing 5 or 7 carbon atoms, for example 2-methylbutyl, n-pentyl, dimethylpropyl , n-hexyl, 2-methylpentyl, 3-methylpentyl, heptyl and the like;

-C _3-7 -cycloalkyl includes cyclic hydrocarbon groups containing 3 to 7 carbons such as cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl and the like .

-C _1-7 -alkoxy is also a straight or branched chain alkoxy having 1 to 7 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, secondary butoxy, 3 Tertiary butoxy, petyloxy, hexyloxy and the like.

In the above compounds, unless specifically noted, preferred substituents are halogen (eg chlorine, bromine, fluorine and iodine), preferably alkyl, alkoxy, aralkyl, or haloalkyl (eg fluoromethyl, Difluoromethyl, trifluoromethyl, 2,2,2-trifluoroethyl, 2,2,3,3,3, -pentafluoropropyl, etc.), nitro, amino, alkylamino, cyano , Formyl, acyl, aminoalkyl, mono- or dialkylaminoalkyl, azide, carboxy, alkoxycarbonyl, carbamoyl, alkylcarbamoyl, azide and the like.

The present invention also includes pharmaceutically acceptable salts, or solvates or hydrates thereof, which the compounds can form.

Pharmaceutically acceptable addition salts include pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. Pharmaceutically acceptable acid addition salts referred to herein include the therapeutically active non-toxic acid addition salt forms that the compounds of the present invention can form. Compounds of the invention having basic properties can be converted to their pharmaceutically acceptable acid addition salts by treating the base form as a suitable acid. Suitable acids are, for example, halosomic acids such as hydrochloric acid or bromic acid; Sulfuric acid; nitric acid; Inorganic acids such as phosphoric acid or acetic acid, trifluoroacetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid (ie, butanedioic acid), maleic acid, fumaric acid, malic acid, tartaric acid, citric acid And organic acids such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclic acid, salicylic acid, p-amino-salicylic acid and pamoic acid.

Compounds of the invention having acidic properties can be converted to their pharmaceutically acceptable base addition salts by treating the acidic form with a suitable organic or inorganic base. Suitable base salt forms include, for example, organic bases such as ammonium salts, alkali and alkaline earth metal salts, such as lithium, sodium, potassium, magnesium, calcium salts and the like, benzatin, N-methyl-D-glucamine, hydrabamine salts. Salts and salts with amino acids such as arginine, lysine and the like.

The present invention also includes hydrates and solvates that the compounds of the present invention can form. The hydrate and solvate can be formed by conventional methods.

Hereinafter, the present invention will be described in more detail.

In the present invention, a small molecule organic compound that directly activates RSK1 was devised based on the world's first discovery that the cells exhibit the effect of promoting cell regeneration and inhibiting expression of transforming growth factors. In fact, according to the experimental example, the direct increase of RSK1 activity by a representative small molecule organic compound selectively phosphorylated the serine No. 105 residue of C / EBPβ, which is a key factor of cell regeneration, and thus activated C / EBPβ inhibited the activity of the target gene. By increasing or inhibiting it increases the therapeutic efficacy of cirrhosis.

The technology according to the present invention is an advanced technology for treating hepatic fibrosis and cirrhosis as well as the inhibition of the conversion growth factor as well as the effect of hepatocyte regeneration. Can also be used as. Furthermore, unlike the specific gene deletion technology, the activity of RSK1 is increased by small molecule organic compounds. Therefore, when the administration of small molecule organic compounds is stopped, the activity of RSK1 can be lowered to a desired level. It becomes an adjustable technology.

In addition, the composition of the present invention can be administered by formulating in a unit dosage form and injection suitable for oral administration according to a conventional method in the pharmaceutical field. Oral dosage forms suitable for this purpose include hard and soft capsules, tablets, powders, suspensions, syrups and the like. Such oral preparations include, in addition to two or more pharmacologically active ingredients, one or more pharmaceutically inert conventional carriers such as excipients such as starch, lactose, carboxymethylcellulose, kaolin, water, gelatin, alcohols, Additional additive components may be included such as binders of glucose, gum arabic, tragacanta rubber, disintegrants such as starch, dextrin, sodium alginate, and lubricants such as talc, stearic acid, magnesium stearate, liquid paraffin, and the like. . In the present invention, a dissolution aid for dissolution may also be added.

The daily dosage of the composition of the present invention depends on a variety of factors, such as the degree of disease progression, time of onset, age, health condition, complications, etc., but is based on the adult in general composition 1 to 500 mg / kg, preferably 30 to 200 mg / kg may be administered several times daily.

The present invention is explained in more detail by the following experimental examples, but the present invention is not limited by these examples.

Experimental Example

Experimental Example 1 <Effect of increasing activity of RSK1 by small molecule organic compounds>

The HKIIE cells, which were cultured for a certain period of time, were incubated with 30 μM of oltipraz, a representative small-molecule organic compound, for 10 minutes-3 hours, and then measured for RSK1 activity in cells. In other words, RSK1 was separated by immunoprecipitation after a certain time after treatment of small molecule organic compound oltipraz, and the enzyme substrate S6 protein and ³² P-ATP were added to the isolated RSK1 and reacted for 30 minutes to S6 protein. Isotope activity contained was measured. According to the experimental results, the activity of RSK1 was continuously increased after 30 minutes of incubation with oltipraz, a small molecule organic compound (FIG. 1A). In addition, the activation of RSK1 in cells was confirmed from the continuous increase in phosphorylation activity of serine residue 380 of the RSK1 molecule in proportion to the increase of RSK1 activity by a typical small molecule organic compound oltipraz (FIG. 1B). Furthermore, oltipraz, a small-molecule organic compound, reconfirmed RSK1 activation by small-molecule organic compounds in cells from migrating RSK1 in the cytoplasm into the nucleus after 6 hours (FIG. 1C).

Experimental Example 2 <In vitro Activity of RSK1 Directly Increased by Oltipraz, a Small Molecule Organic Compound>

To test whether the small molecule organic compound can directly increase the activity of RSK1 even in the state of cell crushing, immunoprecipitation of RSK1 present in the crushed cell was carried out, and then the small molecule organic compound Oltipraz was added at a concentration of 0.01-10 μM. The change in activity of RSK1 was measured by the amount of isotope contained in the S6 protein as a substrate. The kinase enzyme activity of RSK1 in vitro was found to increase significantly from the concentration of 1 μM oltipraz, a small molecule organic compound (FIG. 2A). In the same manner, when the oltipraz was added to immunoprecipitated RSK1 and incubated for 10 minutes-3 hours, the amount of ³² P contained in the S6 protein increased time-dependently (FIG. 2B). This is the first experimental finding that small molecule organic compounds can directly increase the kinase activity of RSK1.

Experimental Example 3 <Virtual Bonding Model of Small-Molecular Organic Compound and RSK1>

It can be hypothesized that oltipraz, an organic compound containing 1,2-dithiole-3-thione and a bicyclic ring structure, directly binds to RSK1 because it directly increases the kinase activity of RSK1. Figure 3 shows one of the possible models of binding between the 725-729 sequence of the RSK1 protein and the dichiol-based compound.

As another small-molecule organic compound, the thiophene derivative is also shown to be able to bind directly to RSK1 as with oltipraz (FIG. 4A). It was also tested whether thiophene derivatives could directly activate RSK1. Kinase activation was measured from the amount of isotope contained in the substrate S6 protein after immunoprecipitating RSK1, and it was observed that RSK1 was directly activated at a concentration of 1 μM of the thiophene derivative (FIG. 4B). These results indicate that the small molecule organic compound or derivative thereof containing 1,2-dithiole-3-thione and its parent nucleus of bicyclic structures (e.g. pyrazine, pyridazine, pyrimidine, thiazole, chiphene, etc.) can be directly incorporated into RSK1. It can be activated with.

Experimental Example 4 <Selective Phosphorylation of C / EBPβ Serine No. 105 Residue by Direct Enhancement of RSK1 by Small Molecule Organic Compound>

Deletion of the C / EBPβ gene in animals prevents liver tissue regeneration (Greenbaum et al., J Clin Invest. 1998; 102 (5): 996-1007), and activation of C / EBPβ is a mediator of liver fibrosis. Inhibits the transforming growth factor (TGF-β) gene expression (Kang et al., FASEB J. 2002; 16 (14): 1988-90). Therefore, activation of C / EBβ plays a key role in the regeneration of liver tissue and the inhibition of hepatic fibrosis. C / EBPβ activation occurs through the phosphorylation of specific amino acid residues of C / EBPβ, namely serine 105 (Buck et al., Mol Cell. 1999; 4 (6): 1087-92). Against this background, we observed whether direct increase of RSK1 activity by small molecule organic compounds induces serine 105 phosphorylation of C / EBβ. Serine 105 phosphorylation of C / EBβ was increased after 3 hours after incubation with oltipraz incubated for a certain period of time, and the amount of pC / EBPβ increased in the cytosolic fraction by the small-molecule organic compound oltipraz. (Ser105) moved to the nuclear fraction after 6 hours (Fig. 5A). In contrast, the degree of phosphorylation of threonine residue No. 188 of C / EBPβ did not change at all, indicating that phosphorylation of C / EBPβ by RSK1 activation by small-molecule organic compounds selectively occurs at specific residues of C / EBPβ molecules.

Phosphorylated C / EBPβ migrated into the nucleus binds to C / EBP binding sites in the promoter region of the target gene to regulate gene expression (Mink et al., Biochem Biophys Acta. 1999; 1447 (2-3): 175-84). Gel delay analysis was performed to test whether the C / EBPβ molecules activated by the small-molecule organic compound Oltipraz and introduced into the nucleus bind to DNA. Enhancement of the binding between the activated C / EBPβ molecule and DNA was confirmed by gel delay analysis, and the gel of the C / EBP-DNA conjugate when anti-pC / EBPβ (S105) antibody was added to the bound C / EBPβ molecule and DNA complex. The movement was further delayed (supershifted) (FIG. 5B). This result demonstrates that p-C / EBPβ (Ser105) is an active C / EBP capable of binding to DNA increased in the nucleus by small molecule organic compounds.

Furthermore, when C / EBPβ expression was observed in whole cell lysates incubated with small-molecule organic compound Oltipraz for 3 to 48 hours, the amount of other C / EBP isoforms did not change significantly but Expression of EBPβ increased significantly with incubation time (FIG. 5C). These results are interpreted as the result of the active ser105-p-C / EBPβ introduced into the nucleus binds to the C / EBP binding site of the C / EBPβ gene promoter to selectively increase the expression of C / EBPβ itself.

In order to directly prove whether phosphorylation of Ser105-pC / EBPβ by small molecule organic compounds is due to RSK1 activity, the cells are overexpressed with RSK1 variants, ie CTT-RSK1 or K112 / 464R-RSK1, which have RSK1 activity inhibitory activity. Was incubated with Oltipraz to measure the amount of Ser105-pC / EBPβ present in whole cell fractions and intranuclear fractions (FIG. 5D). Whole cell fractionation of Ser105-p-C / EBPβ and quantitative increase in nuclei by Oltipraz were completely inhibited by overexpressing a variant of RSK1. However, phosphorylation of C / EBPβ Threonine 188 did not change at all. These results demonstrate that active RSK1 selectively phosphorylates the 105 serine residue of C / EBPβ and demonstrates that phosphorylation of C / EBPβ Ser105 can be promoted by RSK1 activation by small molecule organic compounds.

Luciferase gene expression was measured to observe whether transcriptional activation accompanying the phosphorylation of phosphorylated C / EBPβ and DNA is extinguished by overexpression of RSK1 variants with inhibitory activity of RSK1 (FIG. 5E). Luciferase expression of the target gene (pGL-1651 GST promoter), which is increased by the small molecule organic compound oltipraz, is completely inhibited by overexpression of intracellular RSK1 variants, which is accompanied by increased DNA binding of pC / EBPβ (Ser105). Gene transcriptional activation has been shown to be due to RSK1 activation by Oltipraz.

Experimental Example 5 <Inhibition of TGFβ Gene Expression by Small Molecule Organic Compound>

Transforming growth factor β (TGFβ) is a key mediator of liver fibrosis and cirrhosis. Since the promoter of transforming growth factor contains a C / EBPβ binding site sequence, the effect of RSK1 activator on the expression of transforming growth factor was measured. After overexpressing the transforming growth promoter-luciferase reporter vector, the effect of the small molecule organic compound oltipase on the transcriptional activation ability of the transforming growth promoter after the overexpression of the luciferase enzyme was determined. Transcriptional activity of the transforming growth factor was inhibited in the promoter region of -323 bp to -175 bp containing the sequence (FIG. 6). This result suggests that direct kinase activation of RSK1 by small molecule organic compounds improves liver fibrosis and liver cirrhosis by inhibiting the expression of transforming growth factor (β) through selective phosphorylation of the 105 serine residue of C / EBPβ. Or treatable.

Experimental Example 6 <GSTA Induction Expression by C / EBPβ Activation and C / EBPβ Activation by Newly Synthesized Oltipraz Derivatives>

 As described above, RSK1 activation by oltipraz causes S105 phosphorylation of C / EBβ and expresses a series of genes regulated by C / EBP in the cell. Therefore, the present inventors searched for the increase of Glutathione S-transferase A (GSTA), which is a representative gene induced by C / EBβ activation and C / EBβ activation, using a number of newly synthesized organic compounds, which are oltipraz derivatives. First, in order to select a compound that does not kill cells among the newly synthesized compounds, each synthesized compound (30 uM) was added to the hepatocyte cell and incubated for 24 hours, followed by 4 hours with 0.7 mg / ml of yellow tetrazolium salt (MTT). Further incubation. Since MTT is reduced to formazan crystals which are insoluble in water, the resulting crystals were dissolved in DMSO solvent and absorbance was measured at 595 nm. Newly synthesized compounds (30 uM each), except for compounds which kill cells (marked with * in Table 7), were cultured with the cells for 24 hours and then treated with C / C in nuclear or lysates obtained from treated cells. EBPβ activation and GSTA expression were measured by the aforementioned immunochemical methods. The degree of C / EBPβ activation was observed by C / EBPβ expression, C / EBPβ nuclear migration or C / EBPβ phosphorylation as mentioned in the previous experiment. About 50 new compounds were found to cause C / EBPβ activation as summarized in Table 2 (+, similar to the oltipraz activity potency; ++, stronger than the oltipraz activity potency). These new compounds that cause C / EBPβ activation have been shown to enhance GSTA expression by C / EBPβ activation (o, enzyme induction equivalent to oltipraz; oo, enzyme induction stronger than oltipraz) (Table 2). ). The chemical names of the novel compounds utilized in the examples are summarized in FIG. 8 and the synthesis of representative compounds is described in the following examples.

Example 1

Synthesis of 5- (6-methoxypyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione (CJ-12064)

260 mg (1.00 mmol) of 5- (6-chloropyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione and 0.21 g (0.2 mmol) of sodium carbonate were added to 30 ml of methanol and heated for 4 hours. It was refluxed. After the reaction solution was cooled to room temperature, 10 ml of water and 30 ml of ethyl acetate were added to separate the organic layer. Water was removed with anhydrous sodium sulfate, filtered and the residue obtained by concentration under reduced pressure was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/4) to give 0.18 g of the target compound. 70%)

1H NMR (400MHz, CDCl3) 2.5 (s, 3H), 3.8 (s, 3H), 8.3 (s, 1H), 8.5 (s, 1H)

Mass (FAB) 256 (M +)

Example 2

Synthesis of 5- (6-ethoxypyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione (CJ-12073)

Dissolve 200 mg (0.77 mmol) of 5- (6-chloropyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione in 10 ml of ethanol and add 320 mg (2.32 mmol) of potassium carbonate for 2 hours. It was heated to reflux. After cooling to room temperature, the solvent was removed under reduced pressure and neutralized by adding 25 ml of saturated aqueous ammonium chloride solution. After diluting with 25 ml of dichloromethane, the organic layer was washed with brine and dried over anhydrous sodium sulfate. After removal of the solvent under reduced pressure, the residue was triturated with normal hexane to give 140 mg of the desired compound as a solid (67.5%).

1H NMR (400MHz, CDCl3) 1.45 (t, J = 7Hz, 3H), 2.55 (s, 3H), 4.45 (q, J = 7Hz, 2H), 8.35 (s, 1H), 8.55 (s, 1H)

Mass (EI) 269.8 (M +)

Example 3

Synthesis of 5- (6-isobutoxypyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione (CJ-12074)

Dissolve 200 mg (0.77 mmol) of 5- (6-chloropyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione in 10 ml of isobutanol and add 320 mg (2.32 mmol) of potassium carbonate for 12 hours. Heated to reflux. After cooling to room temperature, 25 ml of saturated aqueous ammonium chloride solution was added and neutralized. 25 ml of ethyl acetate was added for extraction, and the organic solution layer was washed once with water and brine, and then dried over anhydrous sodium sulfate. After removal of solvent under reduced pressure, the residue was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/10) to give 130 mg of the target compound as a solid (56.8%).

1H NMR (400MHz, CDCl3) 1.05 (d, J = 6Hz, 6H), 2.55 (s, 3H), 4.15 (d, J = 6Hz, 1H), 8.35 (s, 1H), 8.55 (s, 1H)

Mass (FAB) 298.7 (M + 1)

Example 4

Synthesis of 5- (6- (2-hydroxyethylthio) pyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione (CJ-12075)

200 mg (0.77 mmol) of 5- (6-chloropyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione was dissolved in 20 ml of tetrahydrofuran and cooled to 0 ° C., followed by 2-mer 0.54 ml (7.70 mmol) of captoethanol and 172 mg (1.53 mmol) of potassium t-butoxide were added sequentially and stirred at the same temperature for 30 minutes. After completion of the reaction by further stirring at room temperature for 2.5 hours, the solvent was removed under reduced pressure, and the residue was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/2) to obtain 170 mg of target. The compound was obtained as a solid. (73.3%)

1H NMR (400MHz, CDCl3) 2.55 (s, 3H), 3.45 (t, J = 5Hz, 2H), 3.95 (t, J = 5Hz, 2H), 8.60 (s, 1H), 8.65 (s, 1H)

Mass (FAB) 303 (M + 1)

Example 5

Synthesis of 5- (6- (3-hydroxypropoxy) pyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione (CJ-12076)

200 mg (0.77 mmol) of 5- (6-chloropyrazin-2-yl) -4-methyl-1,2-dithiol-3-thione was dissolved in 20 ml of tetrahydrofuran and cooled to 0 ° C., followed by 1,3 0.55 ml (7.70 mmol) of propanediol and 172 mg (1.53 mmol) of potassium t-butoxide were added sequentially and stirred at the same temperature for 30 minutes. After completion of the reaction by further stirring at room temperature for 2.5 hours, the solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/1) to obtain 160 mg of the target. The compound was obtained as a solid. (69.4%)

1H NMR (400MHz, CDCl3) 2.10 (q, H = 5Hz, 2H), 2.55 (s, 3H), 3.85 (t, H = 5Hz, 2H), 4.55 (t, H = 5Hz, 2H), 8.35 (s , 1H), 8.55 (s, 1H)

Mass (FAB) 301 (M + 1)

Example 6

Synthesis of 5- (3,4-dichlorophenyl) -1,2-dithiol-3-thione (CJ-12094)

2.24 g (20 mmol) of potassium t-butoxide was added to 50 ml of tetrahydrofuran and suspended at room temperature. 1.89 g (10 mmol) of 3 ', 4'-dichloroacetophenone was diluted in 5 ml of tetrahydrofuran, and slowly added dropwise to the reaction solution, and stirred at the same temperature for 30 minutes. 0.76 g (10 mmol) of carbon disulfide was added and stirred for 30 minutes. Then, 2.67 g (15 mmol) of hexamethyldisilathion was added and stirred for 30 minutes. Then, 3.54 g (15 mmol) of hexachloroethane was added at room temperature to tetrahydrofuran. It melt | dissolved in 10 ml, and stirred for 1 hour. 50 ml of water was added to the reaction solution, and the reaction solution was concentrated under reduced pressure to make the reaction solution about 60 ml, and 50 ml of ethyl acetate was added thereto. The organic layer was separated, dried with anhydrous magnesium sulfate, and then concentrated under reduced pressure. The obtained residue was purified by column chromatography using silica gel (eluent: ethyl acetate / normal hexane = 1/9) to obtain 2.1 g of the target compound (75%).

1H NMR (400MHz, CDCl3) 7.4 (s, 1H), 7.5 (m, 1H), 7.65 (m, 1H), 7.8 (m, 1H)

Mass (FAB) 279 (M +)

Example 7

Synthesis of 5- (2,5-dimethylthiophen-3-yl) -1,2-dithiol-3-thione (CJ-12113)

2.24 g (20 mmol) of potassium t-butoxide was added to 50 ml of tetrahydrofuran and suspended at room temperature. 1.54 g (10 mmol) of 3-acetyl-2,5-dimethylthiophene was diluted in 5 ml of tetrahydrofuran and slowly added dropwise to the reactor and stirred for 30 minutes at the same temperature. 0.76 g (10 mmol) of carbon disulfide was added and stirred for 30 minutes. Then, 2.67 g (15 mmol) of hexamethyldisilathion was added and stirred for 30 minutes, and 3.54 g (15 mmol) of hexachloroethane was added at room temperature for 10 ml of tetrahydrofuran. It was dissolved in and added dropwise and stirred for 1 hour. 50 ml of water was added to the reaction solution to terminate the reaction. The reaction solution was concentrated under reduced pressure to about 60 ml, and 50 ml of ethyl acetate was added thereto. The organic layer was separated, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue obtained was purified by column chromatography using silica gel (eluent: ethyl acetate / normal hexane = 1/9) to obtain 1.7 g of the target compound. (70%)

1H NMR (400MHz, CDCl3) 2.40 (s, 3H), 2.60 (s, 3H), 6.80 (s, 1H), 7.25 (s, 1H)

Mass (FAB) 244.9 (M +)

Example 8

Synthesis of 5-benzo [b] thiophen-2-yl-4-ethyl-1,2-dithiol-3-thione (CJ-12114)

4.44 g (10 mmol) of phosphorus sulphide was added to 50 ml of toluene, and the temperature of the solution was raised to the boiling point while stirring. Then, 2-62 (benzo [b] thiophene-2-carbonyl) -butyric acid methyl ester was added to 5 ml of toluene. A solution of (10 mmoles) was slowly added dropwise to the reaction solution. The reaction was completed by refluxing at the same temperature for 4 hours. After cooling the reaction solution to room temperature, 30 ml of distilled water and 20 ml of methanol were added thereto, and 28% ammonia water (approximately 2 ml) was added to adjust the pH to 8.5. The organic layer was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography using silica gel (eluent: ethyl acetate / normal hexane = 1/9) to obtain 2.35 g of the target compound. (80% )

1 H NMR (400 MHz, CDCl 3) 1.2 (t, 3H), 2.9 (q, 2H), 7.4 (m, 2H), 7.7 (s, 1H), 7.9 (m, 2H)

Mass (FAB) 294.9 (M +)

Example 9

Synthesis of 5-ethyl-4-thiophen-3-yl-1,2-dithiol-3-thione (CJ-12115)

4.44 g (10 mmol) of phosphorus sulfide was added to 50 ml of toluene, and the temperature was raised to the boiling point while stirring. Then, 2.12 g (10 mmol) of 3-oxo-2-thiophen-3-yl-pentanoic acid methyl ester was dissolved in 5 ml of toluene. The solution was slowly added dropwise to the reaction solution. The reaction was completed by refluxing at the same temperature for 4 hours. After cooling the reaction solution to room temperature, 30 ml of distilled water and 20 ml of methanol were added thereto, and 28% ammonia water (approximately 2 ml) was added to adjust the pH to 8.5. The organic layer was separated, dried with anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography using silica gel (eluent: ethyl acetate / normal hexane = 1/9) to obtain 1.5 g of the target compound. (62% )

1 H NMR (400 MHz, CDCl 3) 1.3 (t, 3H), 2.8 (q, 2H), 7.1 (dd, 1H), 7.3 (d, 1H), 7.45 (dd, 1H)

Mass (FAB) 244.9 (M +)

Example 10

Synthesis of 5-phenyl-4,5,6,7-tetrahydrobenzo-1,2-dithiol-3-thione (CJ-12116)

1.42 g (12.6 mmol) of potassium t-butoxide were suspended in a mixed solvent of 20 ml of tetrahydrofuran and 4 ml of dimethylformamide, and 1.0 g (5.74 mmol) of 4-phenylcyclohexanone was added thereto, followed by stirring at room temperature for 15 minutes. 0.38 ml (6.31 mmol) of carbon disulfide was added thereto, stirred at room temperature for 15 minutes, and then 1.82 ml (8.63 mmol) of hexamethyldisilatiane was added thereto, followed by stirring for 15 minutes. After the reaction solution was cooled to 0 ° C., 1.49 g (6.29 mmol) of hexachloroethane was added thereto, followed by stirring at the same temperature for 30 minutes. 10 ml of methanol was added to terminate the reaction, followed by stirring at room temperature for 15 minutes and distillation under reduced pressure to remove the solvent. The residue was diluted with 50 ml of water, extracted with 50 ml of dichloromethane, washed once with brine and dried over anhydrous sodium sulfate. The residue obtained by distillation under reduced pressure was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/30) to give 260 mg of the target compound as a solid (33.3%).

1 H NMR (400 MHz, CDCl 3) 1.95-2.05 (m, 1H), 2.20-2.25 (m, 1H), 2.50-2.60 (m, 1H), 2.90-3.15 (m, 4H), 7.15-7.40 (m, 5H )

Mass (FAB) 265.0 (M + 1)

Example 11

Synthesis of 5-methyl-4-thiophen-3-yl-1,2-dithiol-3-thione (CJ-12145)

4.44 g (10 mmol) of phosphorus penta sulfide was added to 50 ml of toluene, and the temperature was raised to the boiling point while stirring. Then, 1.98 g (10 mmol) of 3-oxo-2-thiophen-3-yl-butyric acid methyl ester was dissolved in 5 ml of toluene. The solution was slowly added dropwise. After refluxing at the same temperature for 4 hours to complete the reaction, the mixture was cooled to room temperature, 30 ml of distilled water and 20 ml of methanol were added thereto, and 28% ammonia water (approximately 2 ml) was added to adjust the pH to 8.5. The organic layer was separated, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography using silica gel (eluent: ethyl acetate / normal hexane = 1/9) to obtain 1.7 g of the target compound. (74%)

1H NMR (400MHz, CDCl3) 2.46 (s, 3H), 7.04 (dd, 1H), 7.17 (d, 1H), 7.52 (d, 1H)

Mass (FAB) 230 (M +)

Example 12

Synthesis of 5- (3-methylthiophen-2-yl) -1,2-dithiol-3-thione (CJ-12146)

2.24 g (20 mmol) of potassium t-butoxide was added to 50 ml of tetrahydrofuran and suspended at room temperature. Then, 1.4 g (10 mmol) of 3-methyl-2-acetylthiophene was diluted in 5 ml of tetrahydrofuran and slowly added dropwise to the reactor. Stir for 30 minutes. 0.76 g (10 mmol) of carbon disulfide was added at the same temperature, followed by stirring for 30 minutes. Then, 2.67 g (15 mmol) of hexamethyldisilatian was added for 30 minutes, followed by stirring for 30 minutes. It was dissolved in and added dropwise, and stirred for 1 hour. 50 ml of water was added to the reaction mixture, and the reaction mixture was concentrated under reduced pressure to make the reaction solution about 60 ml, and 50 ml of ethyl acetate was added thereto. The organic layer was separated, dried over anhydrous magnesium sulfate, and then concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/9) to obtain 1.77 g of the target compound. 77%)

1 H NMR (400 MHz, CDCl 3) 2.4 (s, 3H), 6.8 (s, 1H), 7.2 (m, 1H), 7.3 (m, 1H)

Mass (FAB) 230 (M +)

Example 13

Synthesis of 5- (6-chloropyridin-3-yl) -4-ethyl-1,2-dithiol-3-thione (CJ-12151)

2.24 g (20 mmol) of potassium t-butoxide was added to 50 ml of tetrahydrofuran and suspended at room temperature. Then, 1.83 g (10 mmol) of 1- (6-chloropyridin-3-yl) -butan-1-one was added to tetrahydrofuran. Dilution with 5ml was slowly added dropwise to the reaction solution and stirred at room temperature for 30 minutes. 0.76 g (10 mmol) of carbon disulfide was added and stirred at the same temperature for 30 minutes. Then, hexamethyldisilathion (2.67 g, 15 mmol) was added and stirred for 30 minutes. Then, 3.54 g (15 mmol) of hexachloroethane was added to tetrahydrofuran. It was dissolved in 10 ml dropwise and stirred at room temperature for 1 hour. 50 ml of water was added to the reaction solution to terminate the reaction, and the reaction solution was concentrated under reduced pressure to about 60 ml, and 50 ml of ethyl acetate was added thereto. The organic layer was separated, water was removed with anhydrous magnesium sulfate, and the residue obtained by concentration under reduced pressure was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/4) to obtain 1.09 g of the target compound. 40%)

1 H NMR (400 MHz, CDCl 3) 1.1 (t, 3H), 2.7 (q, 2H), 7.6 (m, 1H), 7.8 (m, 1H), 8.6 (m, 1H)

Mass (FAB) 273 (M +)

Example 14

Synthesis of 4,5-dihydronaphtho [1,2-c] -1,2-dithiol-3-thione (CJ-12162)

0.84 g (7.49 mmol) of potassium t-butoxide was suspended in a mixed solvent of 10 ml of tetrahydrofuran and 2 ml of dimethylformamide, and 0.5 g (3.42 mmol) of alpha-tetralone was added thereto, followed by stirring at room temperature for 15 minutes. 0.23 ml (3.82 mmol) of carbon disulfide was added thereto, followed by stirring at room temperature for 15 minutes, and then 1.1 ml (5.21 mmol) of hexamethyldisilatiane was added thereto, followed by stirring for 15 minutes. After cooling the reaction solution to 0 ° C., 0.89 g (3.76 mmol) of hexachloroethane was added thereto, followed by stirring at the same temperature for 30 minutes. 5 ml of methanol was added to terminate the reaction, followed by stirring at room temperature for 15 minutes, and distillation under reduced pressure to remove the solvent. The residue was diluted with 25 ml of water, extracted with 25 ml of dichloromethane, washed once with brine and dried over anhydrous sodium sulfate. The residue obtained by distillation under reduced pressure was purified by column chromatography on silica gel (eluent: ethyl acetate / normal hexane = 1/60) to give 310 mg of the target compound as a solid (38.3%).

1 H NMR (400 MHz, CDCl 3) 2.90-3.00 (m, 2H), 3.00-3.10 (m, 2H), 7.30-7.35 (m, 2H), 7.45 (m, 1H), 7.65 (m, 1H)

Formulation Example 1

CJ-11781 100mg

Lactose 50mg

Starch 10mg

Magnesium stearate proper amount

The tablets were prepared by mixing the above components and tableting according to a conventional method for producing tablets.

Formulation Example 2

CJ-11795 250mg

Lactose 50mg

Starch 10mg

Magnesium stearate proper amount

The above components were mixed and compressed into tablets according to a conventional method for preparing starch.

Formulation Example 3

CJ-11793 25mg

Lactose 30mg

Starch 28mg

Talc 2mg

Magnesium stearate proper amount

The above ingredients were mixed and filled into gelatin light capsules according to the method for preparing capsules in the usual manner to prepare capsules.

Formulation Example 4

CJ-11808 250mg

10 g of isomerized sugar

30 mg of sugar

Sodium CMC 100mg

Lemon flavor

100ml total by adding purified water

Suspending agent was prepared according to the conventional method for preparing a suspending agent, and filled into a 100 ml brown bottle and sterilized to prepare a suspending agent.

Formulation Example 5

CJ-11759 500mg

20 g of isomerized sugar

20 g of sugar

Sodium Alginate 100mg

Orange flavor

100ml total by adding purified water

According to the conventional method for preparing a suspending agent, a suspending agent was prepared, and a suspending agent was prepared by filling a 100 ml brown bottle and sterilizing it.

Formulation Example 6

CJ-11794 250mg

Lactose 30mg

Starch 20mg

Magnesium stearate proper amount

The above ingredients were mixed closely and filled and sealed in a polyethylin coated cloth to prepare a powder.

Formulation Example 7

Content in 1 tablet of soft capsule

CJ-11843 100mg

Polyethylene Glycol 400 400mg

Concentrated glycerin 55mg

Purified water 35mg

After mixing polyethylene glycol and concentrated glycerin, purified water was added, and the mixture was kept at about 60 ° C, followed by oltifrase, and then mixed uniformly with stirring at about 1,500 rpm with a stirrer. Bubbles were removed using a pump to obtain the contents of the soft capsule. Soft capsule coating is generally prescribed soft gelling agent of gelatin, plasticizer, gelatin 132mg per 1 capsule, concentrated glycerin 52mg, dissolbitol solution 70% 6mg and the amount of ethyl vanillin as a flavor, using carnauba wax as a coating agent It prepared by the usual preparation method.

Increasing the activity of RSK1 directly by the small molecule organic compound according to the present invention not only has the effect of inhibiting the gene expression of the conversion growth factor, but also has a unique pharmacological effect at the molecular level, so as to treat liver cirrhosis as well as regeneration of liver tissue. Organic compounds that activate RSK1 can be used for the prevention and treatment of liver fibrosis and cirrhosis.

Claims (4)

p90 ribosomal S6 kinase 1 (RSK1) A pharmaceutical composition for treating liver fibrosis and cirrhosis, comprising a direct activator. The method according to claim 1, wherein the activator of RSK1 is 5- (2-pyrazinyl) -4-methyl-1,2-dithiol-3-thione (oltipraz) or a compound shown in the following table, a pharmaceutical thereof Pharmaceutical composition for the prevention or treatment of liver fibrosis and cirrhosis, including an acceptable salt thereof, or solvates or hydrates thereof as an active ingredient:

In the above formula, the definition of the symbol is the same as that described in the specification. The pharmaceutical composition for preventing or treating liver fibrosis and cirrhosis according to claim 2, wherein the compound comprises a compound shown in the following table or a pharmaceutically acceptable salt thereof, or a solvate or hydrate thereof as an active ingredient.

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p90 A method for screening substances for the prevention or treatment of liver fibrosis and cirrhosis by measuring the degree of activation of ribosomal S6 kinase 1 (RSK1).

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