The present invention relates to noble sesterterpene compounds and preventive efficacy of the sesterterpene compounds to prevent and treat diabetes, obesity, fatty liver diseases (alcoholic, non-alcoholic, and viral fatty liver diseases), cardiovascular diseases (hyperlipidemia, atherosclerosis) and brain diseases (Parkinson's and Alzheimer's disease). The sesterterpene compounds of the present invention may be used as an effective component of a composition for preventing and treating Type 2 diabetes and secondary diseases caused by diabetes (renal failure and foot ulcer) by controlling expressions of proteins having antidiabetic efficacy and intercellular signals. In addition, the sesterterpene compounds of the present invention may be used as an effective component of a composition for preventing and treating alcoholic, non-alcoholic, and viral fatty liver diseases by inhibiting the formation of fatty acids in the liver and promoting beta-oxidation by which the fatty acids are burn to release the heat to thereby significantly reduce fat accumulation in the liver. In addition, the sesterterpene compounds of the present invention may be used as an effective component of a composition for preventing and treating obesity by inhibiting the differentiation of adipocytes. Moreover, the sesterterpene compounds of the present invention may be used as an effective component of a composition for preventing and treating cardiovascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, alone or in a complex agent together with an agonist of the existing nucleus receptor LXR, by inhibiting activity and generation of pro-inflammatory cytokine as an atherosclerosis factor and inhibiting the formation of NO that controls vasoconstriction. Moreover, the sesterterpene compounds of the present invention may be used as an effective component of a composition for preventing and treating central nervous system diseases such as Parkinson's disease, schizophrenia, and manic-depression, by increasing expression and activity of genes that regulate the function of dopaminergic neuronal cells. Moreover, the sesterterpene compounds of the present invention may be used as an agent for preventing and treating Alzheimer's disease in a complex agent together with agonists of the existing nuclear receptor LXR. Therefore, the sesterterpene compounds developed through the present invention may be used as an effective component of a composition for preventing and treating insulin-independent diabetes mellitus, fatty liver diseases (alcoholic, non-alcoholic, and viral), obesity, cardiovascular diseases (hyperlipidemia and atherosclerosis), and brain disorders (Parkinson's disease and Alzheimer's disease).
Changes in modern man lifestyle, such as nutrient excesses and exercise reduction, have resulted in rapid increases in metabolic diseases such as diabetes, fatty acid, obesity, hyperlipidemia, and atherosclerosis, and central nervous system disorders such as Parkinson's disease and Alzheimer's disease.
Diabetes alone is a major life-threatening disease, and is the direct cause of secondary diseases caused by diabetes, that is, blindness due to chronic complications, end-stage renal failure, neurological disease, and foot ulceration. Of these, the most threatening diseases are cerebrovascular diseases and cardiovascular diseases. It has been known that patients with diabetes are 2 times more likely at risk of getting coronary artery disease and about 3 times more likely at risk of getting peripheral vascular diseases as compared with a normal person. It has been that the reasons are hyperglycemia, dyslipidemia, hyperinsulinemia, hypertension, and changes in the blood clotting mechanism.
Nuclear receptors are ligand-dependent transcription factors to regulate physiological functions such as in vivo metabolism, cell differentiation and apoptosis, and development. Of these, NR4A is considered an orphan nuclear receptor since it does not have a binding region with the co-activator and the accurate ligands thereof have been currently established (Wang et al., Nature, 2003, 423, 555-560). Accordingly, the physiological functions of NR4A have been currently found mainly through studies on overexpression of this gene (gain of function) or selective removal of this gene (loss of function). However, since the recent studies reveal that NR4A also is a ligand-dependent transcription factor, NR4A has emerged as a new medicine molecular target for metabolic disease treatment (Zhan et al., Nat. Chem. Biol., 2008, 4, 548-556).
NR4A reduces expression of sterol regulatory element binding protein-1c (SREBP1C) regulating fatty acid synthesis in the liver (Pei et al., Nat. Med., 2006, 12, 1048-1055; Pols T W et. al., Biochem. Biophys. Res. Commun., 2008, 366, 910-916). It increases the expression of glucose transporter associated with glucose absorption and activates the insulin signaling, resulting in increasing the glucose metabolism, in the muscle and liver (Fu et al., J. Biol. Chem., 2007, 282, 31525-31533). In addition, it increases expressions of uncoupling proteins (UCPs) regulating thermogenesis and PGC-1α and β, lipoprotein lipase (LPL), fatty acid binding protein 4 (FABP4), pyruvate dehydrogenase kinase, and isozyme 4 (PDK4), which regulate fatty acid oxidation, to increase fat metabolism (Pearen, M A et al., Endocrinology, 2006, 147, 5217-5227; Weyrich, P et al., Diabetes, 2009, 58, 2788-2796; Pearen, M A et al., Endocrinology, 2007, 149, 2853-2865; Chao, L C et al., Mol. Endocrinol., 2007, 21, 2152-2163). Therefore, NR4A is developed to have superior efficacy, which can be used as a therapeutic agent for diabetes, fatty liver, and obesity.
As a result of experiment using NR4A in the macrophage, expression of the pro-inflammatory cytokine by LPS or ox-LDL was inhibited by NR4A (Peter I. Bonta et al., Arterioscler. Thromb. Vasc. Biol., 2006, 26, 2288-2294). In addition, NR4A inhibited the proliferation of smooth muscle cells by increasing expression of p27Kip1, cell cycle inhibitor protein, in the vascular smooth muscle and inhibited the occurrence of atherosclerosis by reducing expression of pro-inflammatory cytokines such as IL-11, TNFα, and MCP-1 (Peter I. Bonta et al., Circulation, 2010, 121, 2023-2032). Therefore, NR4A is developed to have superior efficacy, which can be used as a therapeutic agent for atherosclerosis.
LXR agonists reduce serum low-density lipoprotein (LDL) cholesterol and increase high-density lipoprotein (HDL) cholesterol and thus can be developed as a therapeutic agent for hyperlipidemia, and reduce expression of cytokine causing an inflammatory reaction to inhibit atherosclerosis (Dai, Inflammation, 2007, 30, 105-117). In addition, they remove beta amyloid, which is a cause of Alzheimer's disease in the brain, to thereby improve Alzheimer's disease (Riddell, Mol Cell Neuroscie, 2007, 34, 621-628). However, even though the existing LXR agonists have efficacy to treat hyperlipidemia, atherosclerosis, and Alzheimer's disease, they cause fatty acid, resulting in severe hepatotoxicity (Barunowski, J. Physiol. Pharmacol., 2008, 59, 31-55; Chisholm, J. Lipid Res. 2003, 44, 2039-2048). The LXR agonist increase lipid synthesis and lipid peroxidation in the liver, to thereby cause alcoholic, non-alcoholic, and viral fatty liver diseases (Hwahng et al, Hepatology, 2009, 49, 1913-1925; Na et al., Hepatology. 2009, 1122-1131). Moreover, the lipid peroxidation by the LXR agonist inhibits functions of mitochondria to accelerate the occurrence of diabetes (Chu, Mol Cell Biol., 2006, 26, 6786-6798; Liang, J. Biol. Chem., 2002, 277, 9520-9528; Barunowski, J Physiol Pharmacol., 2008, 59, 31-55). That is, since the LXR agonist that have been developed until the present time cause complications such as diabetes or fatty liver in the liver, the LXR agonist in the form of a single agent thereof is not appropriate in being used as a therapeutic agent for hyperlipidemia, atherosclerosis, and Alzheimer's disease. Therefore, the compound inhibiting a side effect (fatty liver) of the existing LXR agonist may be used as an effective component of a composition for preventing and treating diseases such as hyperlipidemia, atherosclerosis, and Alzheimer's disease, in a complex agent together with the developed LXR agonists. Accordingly, the present compound may be used as an effective component of a composition for preventing and treating hyperlipidemia, atherosclerosis, and Alzheimer's disease, in a complex agent together with the existing LXR agonists.
Central nervous system disorders are closely associated with death of dopaminergic neuronal cells. Nurr1, which is one kind of NR4A, is a nuclear receptor protein mainly expressed in the dopaminergic neuronal cell in the midbrain, and directly regulates the expression of genes associated with dopamine synthesis (tyrosine hydroxylase) and dopamine transport (dopamine transporter (Sakurada et al., Development 1999, 40174026; Sacchetti et al., J. Neurochem. 2001, 15651572). It was found from the result of analysis of family history of Parkinson's syndrome that mutation of the Nurr1 gene causes Parkinson's syndrome (Le et al., Nat. Genet., 2002, 33, 85-89). In addition, it was proven that the Nurr1-deficient animal model confirmed the deficient of dopaminergic neuronal cells, and thus the Nurr1 gene is pathologically important in nervous diseases such as Parkinson's syndrome and the like (Joseph et al., Proc. Natl. Acad. Sci. U.S.A., 2002, 100, 15619-15624). Therefore, a Nurr1-activating material may be used as an effective component of a composition for preventing and treating central nervous system disorders such as Parkinson's disease, schizophrenia, and manic-depression.
The present inventors developed new sesterterpene compounds as an NR4A agonist, for preventing and treating insulin-independent diabetes, obesity, fatty liver disease, cardiovascular diseases (hyperlipidemia and atherosclerosis), and brain disorders (Parkinson's disease, schizophrenia, and manic-depression), and then completed the present invention. Further, the present inventors developed new sesterterpene compounds of selectively inhibiting activity of the LXR protein in the liver in order to overcome a side effect of the existing LXR activating ligand (fatty liver), and then completed the present invention. Further, the present inventors verified efficacy of the sesterterpene compounds to prevent and treat diabetes, vascular diseases, and brain disorders (Parkinson's disease) in disease animal models, and then completed the present invention.
The present invention has the following objects.
A first object of the present invention is to provide a compound having superior efficacy to prevent, treat, and improve diabetes and diabetes complications (foot ulcer and renal failure).
A second object of the present invention is to provide a compound having superior efficacy to prevent, treat, and improve vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke.
A third object of the present invention is to provide a compound having superior efficacy to prevent, treat, and improve alcoholic, non-alcoholic, and viral fatty liver diseases.
A fourth object of the present invention is to provide a compound having superior efficacy to prevent, treat, and improve obesity by inhibiting the differentiation of adipocytes.
A fifth object of the present invention is to provide a compound having superior efficacy to prevent, treat, and improve central nervous system disorders such as Parkinson's disease, schizophrenia, and manic-depression.
A sixth object of the present invention is to provide a compound having superior activity to Nurr1.
A seventh object of the present invention is to provide a compound having superior activity to LXR.
An eighth object of the present invention is to provide a pharmaceutical composition for preventing and treating diabetes and diabetes complications (foot ulcer and renal failure), vascular diseases (hyperlipidemia, atherosclerosis, and atherosclerotic stroke), fatty liver disease, obesity, or central nervous system disorders (Parkinson's disease, schizophrenia, and manic-depression), containing a compound of Chemical Formula I below as an effective component.
A ninth object of the present invention is to provide a pharmaceutical composition for preventing and treating vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, containing a compound of Chemical Formula I below as an effective, in a complex agent together with the existing LXR agonists.
A tenth object of the present invention is to provide a pharmaceutical composition for preventing and treating Alzheimer's disease, containing a compound of Chemical Formula I below as an effective component, in a complex agent together with the existing LXR agonists.
An eleventh object of the present invention is to provide a composition for functional food and functional beverage for preventing and improving diabetes and diabetes complications (foot ulcer and renal failure), vascular diseases (hyperlipidemia, atherosclerosis, and atherosclerotic stroke), fatty liver disease, obesity, or central nervous system disorders (Parkinson's disease, schizophrenia, and manic-depression), containing a compound of Chemical Formula I below as an effective component.
A twelfth object of the present invention is to provide a composition for functional food and functional beverage for preventing and improving vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, containing a compound of Chemical Formula I below as an effective component, in a complex agent together with the existing LXR agonists.
A thirteenth object of the present invention is to provide a composition for functional food and functional beverage for preventing and improving Alzheimer's disease, containing a compound of Chemical Formula I below as an effective component, in a complex agent together with the existing LXR agonists.
A fourteenth object of the present invention is to provide a composition for functional cosmetics for preventing and improving obesity, containing a compound of Chemical Formula I below as an effective component.
A fifth object of the present invention is to provide a composition for functional feedstuff for preventing and improving diabetes and diabetes complications (foot ulcer and renal failure), vascular diseases (hyperlipidemia, atherosclerosis, and atherosclerotic stroke), fatty liver disease, obesity, or central nervous system disorders (Parkinson's disease, schizophrenia, and manic-depression), containing a compound of Chemical Formula I below as an effective component.
A sixteenth object of the present invention is to provide a composition for functional feedstuff for preventing and improving vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, containing a compound of Chemical Formula I below as an effective component, in a complex agent together with the existing LXR agonists.
A seventeenth object of the present invention is to provide a composition for functional feedstuff for preventing and improving Alzheimer's disease, containing a compound of Chemical Formula I below as an effective component, in a complex agent together with the existing LXR agonists.
An eighteenth object of the present invention is to provide a pharmaceutical composition for preventing and treating diseases through Nurr1 activation.
A nineteenth object of the present invention is to provide a pharmaceutical composition for preventing and treating diseases through LXR inhibitory activity.
A twentieth object of the present invention is to provide a composition for functional food and function beverage for preventing and improving diseases through Nurr1 activation.
A twenty-first object of the present invention is to provide a composition for functional food and functional beverage for preventing and treating diseases through LXR inhibitory activity.
The present invention is directed to a noble sesterterpene compound, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, and uses of these, and the sesterterpene compound provides superior activity to prevent and treat diabetes, foot ulcer and renal failure due to diabetes, alcoholic, non-alcoholic, and viral fatty liver diseases, obesity, vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, or central nervous system disorders such as Parkinson's disease, Alzheimer's disease, schizophrenia, and manic-depression.
The sesterterpene compound of the present invention is represented by Chemical Formula I below:
[in Chemical Formula I,
W is C(=A) or CR11R12, A is O, S, or NR13, and R13 is hydrogen, OH, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C6-C20)aryl or (C4-C20)heteroaryl;
R11 and R12 each are independently hydrogen, -L-R14, halogen, —N3, (C2-C20) heteroaryl, (C6-C20) aryl, —NR15R16,
(C2-C8)alkenyl, or (C2-C8)alkynyl, but both R11 and R12 are not hydrogen;
L is O, NR13, S, SO2, or Se;
R14 is hydrogen, (C1-C8)alkyl, (C1-C8)alkylcarbonyl, (C6-C20) aryl, —SO2R17, —(CH2)m—R8, (C2-C8) alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid, glucose, or
R17 is (C6-C20)aryl, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid, glucose, or
R15 and R16 each are independently hydrogen, (C1-C8)alkyl, —(CH2)m—R18, (C2-C8) alkenyl, (C2-C8) alkynyl, (C3-C8)cycloalkyl, (C6-C20)aryl, (C4-C20)heteroaryl, amino acid, glucose, or
R18 is (C2-C8)alkenyl, (C2-C8)alkynyl, —NR19R20, or (C2-C20)heteroaryl;
X is a single bond, CR20, O, S, or NR20;
R19 and R20 each are independently hydrogen, (C1-C8)alkyl, (C6-C20)aryl, (C2-C8)alkenyl, (C2-C8) alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid, or
R1 is —CH2R21, —COOH, —C(═O)R22, (C2-C20) heteroaryl,
R21 is —OR26, N3, —NR15R16, halogen, CN, NO2, (C2-C20)heteroaryl,
—SR20, —SO2R17, —SeR20, glucose, or amino acid;
R22 to R25 each are independently hydrogen, CN, halogen, (C1-C8)alkyl, OR14,
R26 is hydrogen, (C1-C8)alkyl, (C6-C20)aryl, (C1-C8)alkyldi(C6-C20)arylacetyl, halo(C1-C8)alkyldi(C6-C20)arylacetyl, (C1-C8)alkyldi(C6-C20)arylsilyloxy, —(CH2)mR27, —C(O)R27, —S(═O)2R28, —P(═O) (R28)2, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid,
Y′ is O, S, or NR″; Z′ is a single bond, NH, O, S, or Se; R′ and R″ each are independently hydrogen, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C6-C20)aryl, or (C4-C20)heteroary;
R27 is (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C6-C20)aryl, (C2-C20)heteroaryl, 5- to 6-membered heterocycloalkyl, or
R28 is (C1-C8)alkyl, (C6-C20) aryl, (C1-C8)alkoxy, (C6-C20)aryloxy, (C2-C20)heteroaryl, OH, or OM;
M is alkali metal;
R4 is hydrogen, (C1-C8)alkyl, or -L-R29; R29 is hydrogen, (C1-C8)alkyl, (C1-C8)alkylcarbonyl, (C6-C20)aryl, or —SO2R17;
R6 is hydrogen, (C1-C8)alkyl, or OH;
R2, R3, R5, R7 and R8 each are independently hydrogen, (C1-C8)alkyl, (C6-C20)aryl, or (C2-C20)heteroaryl, and R2 and R3, R5 and R6, and R7 and R8 are independently linked to each other to form a double bond, linked via oxygen (O) to form epoxide, or linked via sulfur (S) to form thiirane;
R9 is (C1-C8)alkyl, CHO, or COOH, and R9 may form a double bond together with R8;
R10 is —(CH2)mR30, CHO, COOH,
R30 is hydrogen, halogen, OH, —NR15R16, SH, or SeH;
R31 is (C1-C8)alkyl,
R32 to R35 each are independently hydrogen, (C1-C8)alkyl, (C6-C20)aryl, —OH, —SH, —NR15R16, —O—OH, amino acid, glucose, or
Z is O or S;
m is an integer of 1 to 5;
the heteroaryl of R1 and R21, the alkyl, aryl, and heteroaryl of R2, R3, R5, R7 and R8, the alkyl of R4, R6, R9, R22 to R25 and R31, the heteroaryl, aryl, alkenyl, and alkynyl of R11 and R12, the alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl of R13, R15, R16, R19, R20, R′ and R″, the alkyl, alkylcarbonyl, aryl, alkenyl, alkynyl, cycloalkyl, and heteroaryl of R14, the aryl and alkyl of R17 and R32 to R35, the alkenyl, alkynyl, and heteroaryl of R18, the alkyl, aryl, alkyldiarylsilyloxy, alkenyl, alkynyl, cycloalkyl, heteroaryl of R26, the alkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocycloalkyl of R27, the alkyl, aryl, alkoxy, aryloxy, and heteroaryl of R28, the alkyl, alkylcarbonyl, and aryl of R29 each may be further substituted with at least one substituent selected from a group consisting of (C1-C8)alkyl, halogen, (C6-C20)aryl, (C6-C20)ar(C1-C8)alkyl, halo(C1-C8)alkyl, cyano, nitro, (C1-C8)alkoxy, (C6-C20)aryloxy, (C1-C8)alkylthio, (C6-C20)arylthio, amino, mono- or di-(C1-C8)alkylamino, mono- or di-(C6-C20)arylamino, (C1-C8)alkyl(C6-C20)arylamino, (C1-C8)alkylcarbonyl, (C6-C20)arylcarbonyl, (C1-C8)alkoxycarbonyl, (C6-C20)aryloxycarbonyl, (C2-C20)heteroaryl, hydroxy, formyl, and carboxyl; and
the heteroaryl and heterocycloalkyl contains at least one hetero atom selected from N, O, and S.]
Specific examples of the compound of Chemical Formula I are as follows:
[Y is O or S;
W is
R1 is
D is amino acid;
R4 is H, CH3, OH, SH, SeH, OTs, OMe, OAc, C(CN)2, or
R6 is H or
R9 is CH3, CH2OH, CHO, or COOH;
R10 is CH2CH3, CHO, COOH, CH2Cl, CH2F, CH2NH2, CH2OH, CH2SH, CH2SeH,
The sesterterpene compound of Chemical Formula I may be represented by Chemical Formula II below, and the present invention provides a novel sesterterpene compound of Chemical Formula II below:
In Chemical Formula II,
W is C(=A) or CR11R12, A is O, S, or NR13, and R13 is hydrogen, OH, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C6-C20)aryl or (C4-C20)heteroaryl;
R11 and R12 each are independently hydrogen, -L-R14, halogen, —N3, (C2-C20) heteroaryl, (C6-C20) aryl, —NR15R16,
(C2-C8)alkenyl, or (C2-C8)alkynyl, but both R11 and R12 are not hydrogen;
L is O, NR13, S, SO2, or Se;
R14 is hydrogen, (C1-C8)alkyl, (C1-C8)alkylcarbonyl, (C6-C20)aryl, —SO2R17, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid, glucose, or —(CH2)m—R18; and R17 is (C6-C20) aryl, (C2-C8) alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid, glucose, or (C1-C8)alkyl;
R15 and R16 each are independently hydrogen, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C6-C20)aryl, (C4-C20)heteroaryl, amino acid, glucose, or —(CH2)m—R18;
R18 is (C2-C8) alkenyl, (C2-C8) alkynyl, —NR19R20, or (C2-C20)heteroaryl;
X is a single bond, CR20, O, S, or NR20; R19 and R20 each are independently hydrogen, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid, or (C6-C20)aryl;
R1 is —CH2R21, —COOH, —C(═O)R22, (C2-C20) heteroaryl,
R21 is —OR26, N3, —NR15R16, halogen, CN, NO2, (C2-C20) heteroaryl,
—SO2R17, —SeR20, glucose, amino acid, or —SR20;
R22 to R25 each are independently hydrogen, CN, halogen, (C1-C8)alkyl, OR14,
R26 is hydrogen, (C1-C8)alkyl, (C6-C20)aryl, (C1-C8)alkyldi(C6-C20)arylacetyl, halo(C1-C8)alkyldi(C6-C20)arylacetyl, (C1-C8)alkyldi(C6-C20)arylsilyloxy, —(CH2)mR27, —C(O)R27, —S(═O)2R28, —P(═O) (R28)2, (C2-C8) alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, (C4-C20)heteroaryl, amino acid,
R27 is (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C6-C20)aryl, (C2-C20)heteroaryl, 5- to 6-membered heterocycloalkyl, or
R28 is (C1-C8)alkyl, (C6-C20) aryl, (C1-C8)alkoxy, (C6-C20)aryloxy, (C2-C20)heteroaryl, OH, or OM;
M is alkali metal;
R4 is hydrogen, (C1-C8)alkyl, or -L-R29; R29 is hydrogen, (C1-C8)alkyl, (C1-C8)alkylcarbonyl, (C6-C20)aryl, or —SO2R17;
R6 is hydrogen, (C1-C8)alkyl, or OH;
R2, R3, R5, R7 and R8 each are independently hydrogen, (C1-C8)alkyl, (C6-C20)aryl, or (C2-C20)heteroaryl, and R2 and R3, R5 and R6, and R7 and R8 are independently linked to each other to form a double bond, linked via oxygen (O) to form epoxide, or linked via sulfur (S) to form thiirane;
R9 is (C1-C8)alkyl, CHO, or COOH;
R10 is —(CH2)mR30, CHO, COOH,
R30 is hydrogen, halogen, OH, —NR15R16, SH, or SeH;
R31 is (C1-C8)alkyl,
R32 to R35 each are independently hydrogen, (C1-C8) alkyl, (C6-C20) aryl, —OH, —SH, —NR15R16, amino acid, glucose, —O—OH, or
Z is O or S;
m is an integer of 1 to 5;
the heteroaryl of R1, R18, and R21, the alkyl, aryl, and heteroaryl of R2, R3, R5, R7, R8 and R28, the alkyl of R4, R6, R9, R15, R16, R22 to R25 and R31, the heteroaryl and aryl of R11 and R12, the alkyl and aryl of R14, R17, R19, R20, R26, R29, and R32 to R35, and the alkyl, aryl, heteroaryl, and heterocycloalkyl of R27 each may be further substituted with at least one substituent selected from a group consisting of (C1-C8)alkyl, halogen, (C6-C20)aryl, (C6-C20)ar(C1-C8)alkyl, halo(C1-C8)alkyl, cyano, nitro, (C1-C8)alkoxy, (C6-C20)aryloxy, (C1-C8)alkylthio, (C6-C20)arylthio, amino, mono- or di-(C1-C8)alkylamino, mono- or di-(C6-C20)arylamino, (C1-C8)alkyl(C6-C20)arylamino, (C1-C8)alkylcarbonyl, (C6-C20)arylcarbonyl, (C1-C8)alkoxycarbonyl, (C6-C20)aryloxycarbonyl, (C2-C20)heteroaryl, hydroxy, formyl, and carboxyl; and
the heteroaryl and heterocycloalkyl contains at least one hetero atom selected from N, O, and S.]
Specific examples of the compound of Chemical Formula II are as follows:
[Y is O or S;
W is
R1 is
R4 is H, CH3, OH, SH, SeH, OTs, OMe, OAc, C(CN)2, or
R6 is H or
R9 is CH3, CH2OH, CHO, or COOH;
R10 is CH2CH3, CHO, COOH, CH2Cl, CH2F, CH2NH2, CH2OH, CH2SH, CH2SeH,
Further, the present invention provides a pharmaceutical composition for preventing and treating diabetes and diabetes complications (foot ulcer or renal failure), the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a pharmaceutical composition for preventing and treating vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a pharmaceutical composition for preventing and treating alcoholic, non-alcoholic, and viral fatty liver diseases, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a pharmaceutical composition for preventing and treating obesity, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a pharmaceutical composition for preventing and treating central nervous system disorders such as Parkinson's disease, schizophrenia, and manic-depression, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a pharmaceutical composition for preventing and treating vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component, in a complex agent together with the existing LXR agonists.
Further, the present invention provides a pharmaceutical composition for preventing and treating Alzheimer's disease, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component, in a complex agent together with the existing LXR agonists.
Further, the present invention provides a pharmaceutical composition for Nurr1 activation, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a pharmaceutical composition for LXR inhibitory activity, the pharmaceutical composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a composition for functional food and beverage for preventing and improving diabetes and diabetes complications (foot ulcer or renal failure), the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive.
Further, the present invention provides a composition for functional food and beverage for preventing and improving vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive.
Further, the present invention provides a composition for functional food and beverage for preventing and improving alcoholic, non-alcoholic, and viral fatty liver diseases, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a composition for functional food, beverage, and cosmetics for preventing and improving obesity, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive.
Further, the present invention provides a composition for functional food and beverage for preventing and improving central nervous system disorders such as Parkinson's disease, schizophrenia, and manic-depression, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a composition for functional food and beverage for preventing and improving vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive, and the existing LXR agonist.
Further, the present invention provides a composition for functional food and beverage for preventing and improving Alzheimer's disease, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive, and the existing LXR agonist.
Further, the present invention provides a composition for functional feedstuff for preventing and improving diabetes and diabetes complications (foot ulcer or renal failure), the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive.
Further, the present invention provides a composition for functional feedstuff for preventing and improving vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive.
Further, the present invention provides a composition for functional feedstuff for preventing and improving alcoholic, non-alcoholic, and viral fatty liver diseases, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a composition for functional feedstuff for preventing and improving obesity, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive.
Further, the present invention provides a composition for functional feedstuff for preventing and improving central nervous system disorders such as Parkinson's disease, schizophrenia, and manic-depression, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a salt thereof acceptable as a food additive, as a pharmaceutically acceptable carrier and an effective component.
Further, the present invention provides a composition for functional food and beverage for preventing and improving diseases through Nurr1 activation, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or an acceptable salt thereof. The diseases include diabetes, diabetes complications (foot ulcer and renal failure), vascular diseases (hyperlipidemia, atherosclerosis, and atherosclerotic stroke), fatty livers (alcoholic, non-alcoholic, or viral fatty liver diseases), obesity, and central nervous system disorders (Parkinson's disease, schizophrenia, and manic-depression).
Further, the present invention provides a composition for functional food and beverage for preventing and improving diseases through LXR inhibitory activity, the composition containing the compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or an acceptable salt thereof. The diseases include diabetes, diabetes complications (foot ulcer and renal failure), vascular diseases (hyperlipidemia, atherosclerosis, and atherosclerotic stroke), fatty livers (alcoholic, non-alcoholic, or viral fatty liver diseases), obesity, and central nervous system disorders (Parkinson's disease, schizophrenia, and manic-depression).
Examples of the LXR agonist mentioned in the present invention are as follows. However, the present invention is not limited to the materials exemplified below, and is applied to all the LXR agonists [the existing LXR agonists: WO2010/054229, WO2010/039529, WO2010/023317, WO2009/150109, WO2009/040289, WO2009/024550, WO2009/021868, WO2008/119657, WO2008/073825, WO2007/092065, WO2007/081335, WO2007/050425, WO2007/050271, WO2007/047991, WO2007/002563, WO2007/002559, WO2006/109633, WO2006/073367, WO2006/073366, WO2006/073365, WO2006/073364, WO2006/073363, WO2006/066779, WO2006/046593, WO2006/037480, WO2006/017384, WO2006/003923, WO2005/121093, WO2005/113499, WO2005/077124, WO2005/077122, WO2005/058834, WO2005/023782, WO2005/023247, WO2005/023196, WO2005/023188, WO2005/016277 WO2005/005417, WO2005/005416, WO2004/076418, WO2004/072041, WO2004/026816, WO2004/024162, WO2004/024161, WO2004/011448, WO2004/009091, WO2003/106435, WO2003/099775, WO2003/099769, WO2003/090869, WO2003/090746, WO2003/090732, WO2003/082802, WO2003/082205, WO2003/082192, WO2003/060078, WO2003/059884, WO2003/059874, WO2003/053352, WO2003/045382, WO2003/031408, WO2002/062302, WO2002/024632, WO2001/060818, WO2001/003705, WO2000/066611, WO2000/054759, WO1997/028137, EP1398032, etc.].
The LXR agonist is preferably N-(2,2,2-trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1-(trifluoromethyl)ethyl]phenyl]-benzenesulfonamide (T0901317) or 3-[3-[[[2-chloro-3-(trifluoromethyl)phenyl]methyl](2,2-diphenylethyl)amino]propoxy]benzeneacetic acid (GW3965).
The amount of the sesterterpene compound of Chemical Formula I, a stereoisomer thereof, an enantiomer thereof, an in vivo-hydrolysable precursor thereof, or a pharmaceutically acceptable salt thereof, which is used to achieve therapeutic effects according to the present invention is varied depending on the specific compound, administration method, subject to be treated, and disease to be treated, but depends on the conventional medicine administration amount. More preferably, the compound of Chemical Formula I may be administered in the range of an effective input amount of 1-100 mg/kg (body weight)/1 day. In addition, the compound is administered one per day or several times per day within the range of an effective input amount. In addition, oral administration or topical administration may be possible depending on the kind of dosage form. The pharmaceutical composition according to the present invention may be formulated into all of the existing various forms in the case of oral administration, and may be in various forms such as tablet, powder, dry syrup, chewable tablet, granule, chewing tablet, capsule, soft capsule, pill, drink, sublingual tablet, and the like. The tablet according to the present invention may be administered to a patient in an effective amount through any bio-available form or manner, that is, an oral pathway. An appropriate form or manner may be easily selected depending on characteristics of a disease state to be treated or prevented, stage of the disease, and other related matter. If the composition according to the present invention is in a tablet form, it may further include at least one pharmaceutically acceptable vehicle, and the ratio of properties of the vehicle may be determined by dissolution and chemical properties of the selected tablet, the selected administration pathway, and the standard pharmaceutical practice.
The sesterterpene compound of Chemical Formula I according to the present invention has superior Nurr1 activation, and thus can control and maintain blood sugar, treat insulin-independent diabetes, and prevent the occurrence of diabetes complications. Further, the sesterterpene compound of Chemical Formula I according to the present invention can be used as an effective component of a composition for improving, treating, and preventing diabetes and diabetes complications (foot ulcer and renal failure) by improving insulin sensitivity through regulation of hormones associated with glucose metabolism and protecting pancreatic function to thereby control fasting blood sugar and prevent the occurrence of diabetes complications (foot ulcer and renal failure).
Further, the sesterterpene compound of Chemical Formula I according to the present invention can have a superior effect in inhibiting differentiation of adipocytes, and thus can be useful in improving, preventing, and treating obesity.
Further, the sesterterpene compound of Chemical Formula I according to the present invention may be used as an effective component of a composition for preventing, treating, and improving alcoholic, non-alcoholic, and viral fatty liver diseases by inhibiting the generation of fatty acids in the liver and promoting beta-oxidation by which the fatty acids are burned to release the heat to thereby significantly reduce fat accumulation in the liver.
Further, the sesterterpene compound of Chemical Formula I according to the present invention reduces low-density lipoprotein (LDL) cholesterol and inhibits expressions of inflammatory cytokines derived from macrophage and endotheliocyte, signaling proteins, and lipid biosynthesis enzymes, and thus can be used as an effective component of a composition for improving, treating, and preventing vascular diseases such as hyperlipidemia and atherosclerosis, alone or in a complex agent together with the existing LXR agonists that have been developed until the present time.
Further, the sesterterpene compound of Chemical Formula I according to the present invention that activates Nurr1 may be used alone as an effective component of a composition for improving, treating, and preventing brain disorders such as Parkinson's disease, schizophrenia, and manic-depression.
Further, the sesterterpene compound of Chemical Formula I according to the present invention may be used as an effective component of a composition for improving, treating, and preventing Alzheimer's disease in a complex agent together with the existing LXR agonists.
Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the embodiments are used to exemplify the present invention. The present invention may be variously modified and changed without being limited by the embodiments.
The present invention has an intent to disclose a compound of Chemical Formula I, stereoisomer thereof, enantiomer thereof, in vivo-hydrolysable precursor thereof, or pharmaceutically acceptable salt thereof, and uses of these materials as an agent for preventing and treating diabetes, foot ulcer and renal failure due to the diabetes, obesity, fatty liver, vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, and brain disorders such as Parkinson's disease and Alzheimer's disease.
The present invention has an intent to disclose a compound of Chemical Formula I, stereoisomer thereof, enantiomer thereof, in vivo-hydrolysable precursor thereof, or pharmaceutically acceptable salt thereof, and uses of these materials as an effective component of functional food and beverage, helpful in preventing and improving diabetes, foot ulcer and renal failure due to the diabetes, obesity, fatty liver, vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, and brain disorders such as Parkinson's disease and Alzheimer's disease.
The present invention has an intent to disclose a compound of Chemical Formula I, stereoisomer thereof, enantiomer thereof, in vivo-hydrolysable precursor thereof, or pharmaceutically acceptable salt thereof, and uses of these materials as an effective component of cosmetics, helpful in preventing and improving obesity.
The present invention has an intent to disclose a compound of Chemical Formula I, stereoisomer thereof, enantiomer thereof, in vivo-hydrolysable precursor thereof, or pharmaceutically acceptable salt thereof, and uses of these materials as an effective component of functional feedstuff, helpful in preventing and improving diabetes, foot ulcer and renal failure due to the diabetes, obesity, fatty liver, vascular diseases such as hyperlipidemia, atherosclerosis, and atherosclerotic stroke, and brain disorders such as Parkinson's disease and Alzheimer's disease.
In addition, the present invention provides a noble sesterterpene compound of Chemical Formula II.
Phorbaketal A (10 mg, 0.025 mmol, Rho et. al., Organic Letters, 2009, 11, 5590-5593) was dissolved in dichloromethane, and then p-TsCl (5.4 mg, 1.2 eq) and triethylamine (0.01 mmol) were put therein, followed by stirring for 5 hours. The reaction was terminated by an aqueous saturated NaHCO3 solution and water (40 ml). The organic solvent layer was washed with water twice, followed by drying over Na2SO4, and then concentration was conducted by using an evaporation concentrator. The resultant material was purified by using silica column chromatography, to obtain Compound 1. MS m/z 554 [M+H]+
Compound 1 (10 mg, 0.018 mmol) was dissolved in dimethylformamide (DMF, 5 ml), and NaN3 (11.7 mg, 10 mmol) was put therein, and then the reaction was allowed to proceed at 70° C. under nitrogen conditions for 8 hours. After the reaction liquid was cooled, ice water was put therein, followed by washing with water, and then the organic solvent layer was separated. After drying over Na2SO4 and distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 2. MS m/z 424 [M+H]+
Compound 2 (10 mg, 0.023 mmol) was dissolved in acetonitril (5 ml), and NaI (0.20 mmol) was put and FeCl3 (0.032 mmol) was put therein. After the reaction liquid was stirred for 20 minutes, the reaction was terminated by adding chloroform (5 ml) thereto. After washing with an aqueous Na2SO3 solution and an aqueous NaHCO3 solution, the obtained organic solvent layer was again washed with salt water. The organic solvent layer was dried over Na2SO4 and distilled under reduced pressure, and then the obtained residue was purified by silica column chromatograph to obtain Compound 3. MS m/z 398 [M+H]+
Phorbaketal A (20 mg, 0.050 mmol) was dissolved in dichloromethane (8 ml), and then Dess-Martin periodinane (21.3 mg, 0.050 mmol) prepared in dichloromethane (10 ml) was added thereto, followed by stirring. After 30 minutes, dichloromethane (50 ml) was put in the homogeneous reaction liquid, and then 1.3M NaOH (20 ml) and water (25 ml) were added thereto. The organic solvent layer was distilled under reduced pressure to obtain a residue, followed by silica column chromatography, to obtain Compound 4 (15.9 mg, 80%). 1H-NMR (300 MHz, CDCl3): d1.59 (3H, s), 1.67 (3H, s), 1.78 (3H, s), 1.79 (3H, s), 1.87 (3H, s), 2.12 (6H, m), 2.38 (1H, t), 2.71 (1H, dd), 3.02 (1H, d), 4.56 (1H, s), 4.82 (1H, t), 5.11 (1H, s), 5.27 (1H, d), 5.40 (1H, s), 6.53 (1H, s), 6.67 (1H, d), 9.52 (1H, s). MS m/z 397 [M+H]+
80% NaClO2 (45.3 mg, 20 eq) and NaH2PO4.2H2O (52 mg, 15 eq.) were prepared in water (5 ml), and then Compound 4 (10 mg, 0.025 mmol) and 2-methylbut-2-ene (10 ml) prepared in tert-butyl alcohol (10 ml) at 0° C. were added thereto. Dioxane (5 ml) was added thereto, and the mixture was stirred at room temperature for 8.5 hours. After the resultant material was diluted with water (45 ml), the target product was extracted with chloroform (2*30 ml) and then washed with salt water (60 ml) and water (40 ml), followed by drying over Na2SO4. The residue left after distillation under reduced pressure was purified by silica column chromatography, to obtain an amorphous type Compound 5 (9.5 mg, 91% yield). 1H-NMR (300 MHz, CDCl3): d1.63 (3H, s), 1.69 (3H, s), 1.79 (3H, s), 1.83 (3H, s), 1.92 (3H, s), 2.03-2.16 (6H, m), 2.36 (1H, t), 2.71 (1H, dd), 2.99 (1H, d), 4.50 (1H, s), 4.76 (1H, t), 5.12 (1H, s), 5.24 (1H, d), 5.35 (1H, s), 6.39 (1H, s), 6.73 (1H, d). MS m/z 413 [M+H]+
Compound 5 (10 mg, 0.024 mmol) was dissolved in anhydrous dichloromethane (5 ml), and then oxalyl chloride (1.0 ml) was put therein, followed by stirring for 3 hours. Excess oxalyl chloride was removed by distillation under reduced pressure, followed by mixing with acid chloride prepared in dichloromethane (2 ml), and then piperidine (2 mg, 0.024 mmol) was added thereto. The mixture was stirred for 1 hour, followed by mixing with water, and then the organic solvent layer was separated and then washed with water (25 ml). After drying over Na2SO4, the obtained residue was purified by column chromatography, to obtain Compound 6 (1.9 mg, 92%). 1H-NMR (300 MHz, CDCl3):d1.56-1.68 (12H, s) 1.76 (6H, s) 1.88 (3H, s) 2.04-2.13 (6H, m) 2.54 (2H, d) 3.92 (1H, m), 3.51 (4H, bs) 4.66 (1H, s) 4.77 (1H, s) 5.10 (1H, s) 5.23 (1H, d) 5.36 (1H, s), 5.63 (1H, s) 6.65 (1H, s). MS m/z 480 [M+H]+
Phorbaketal A (10 mg, 0.025 mmol) was dissolved in dichloromethane (2 ml), and then diethylaminosulfur trifluoride (DAST, Et2NSF3) (6.06 mg, 1.5 eq) prepared in dichloromethane (5 ml) was slowly added thereto at −78° C. The temperature of the reaction liquid was raised to room temperature, and then mixed with water. The organic solvent layer was washed with water, followed by drying over Na2SO4 and distillation under reduced pressure, and the obtained residue was purified by silica gel column chromatography. 7.8 mg of Compound 7 was obtained (78%). 1H-NMR (300 MHz, CDCl3): d1.61 (3H, s), 1.70 (3H, s), 1.77 (3H, s), 1.78 (3H, s), 1.85 (3H, s), 2.08-2.16 (6H, m), 2.512.68 (3H, m), 4.61 (1H, d), 4.71-4.83 (2H, m), 4.92 (1H, m), 5.10 (1H, d), 5.27 (1H, d), 5.35 (1H, s), 5.73 (1H, d), 6.67 (1H, d). MS m/z 401 [M+H]+
Phorbaketal A (20 mg, 0.050 mmol) was prepared in anhydrous DMF (10 ml). Anhydrous K2CO3 (6.93 mg, 2 eq) was added thereto, and the mixture liquid was allowed react at 40° C. for 30 minutes. Propargyl bromide (3-bromopropyne, 12 mg, 2 eq) was slowly added to the mixture liquid, and the reaction was allowed to further proceed for 6 hours while the progress of the reaction was confirmed by TLC. The reaction was terminated by water (50 ml), followed by extraction with ethylacetate (3*50 ml). The organic solvent layer was washed with water (50 ml*2), and then dried over anhydrous Na2SO4. After distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 815.2 mg (70%). MS m/z 437 [M+H]+
Compound 8 (10 mg, 0.022 mmol) and phenylazide (2.7 mg, 0.022 mol) were dissolved in DMF (5 ml), and then 1M sodium ascorbate (0.2 ml, 0.11 mmol) and 1M CuSO4 (0.1 ml, 10 mol %) were sequentially added while stirring at 6500. The reaction solution was stirred at 650 for 24 hours, and the reaction was terminated by slowly adding cold water. The precipitate generated due to addition of water was filtered, and then washed with water, followed by purification by silica column chromatography. MS m/z 556 [M+H]+
Compound 2 (10 mg, 0.023 mmol) and phenylacetylene (2.4 mg, 0.023 mmol) were prepared in DMF (5 m), and then 1M sodium ascorbate (0.2 ml, 0.11 mmol) and IM CuSO4 (0.1 ml, 10 mol %) were sequentially added while stirring at 65° C. The reaction liquid was stirred at 65° C. for 24 hours, and the reaction was terminated by slowly adding cold water. The precipitate generated due to addition of water was filtered, and then washed with water, followed by purification by silica column chromatography. MS m/z 526 [M+H]+
Phorbaketal A (8 mg, 0.017 mmol) was mixed with methanol (4 ml), and K2CO3 (3.6 mg, 0.026 mmol) was added thereto, and then the mixture was stirred at room temperature for 2 hours. The reaction liquid was layer-separated by using water (50 ml) and ethylacetate (3*50 ml). The organic solvent layer was dried over Na2SO4 and then distilled under reduced pressure, and the obtained residue was purified by column chromatography, to obtain Compound 11. MS m/z 415 [M+H]+
Phorbaketal A (10 mg, 0.024 mmol) was dissolved in dichloromethane (5 ml), and then m-CPBA (10.2 mg, 2.4 eq) dissolved in dichloromethane (6 ml) was slowly mixed therewith at 0° C. After stirring at room temperature for 3 hours, an aqueous saturated NaHCO3 solution was put therein, followed by further stirring for 30 minutes. The reaction mixture was extracted with dichloromethane, and the organic solvent layer was washed with water, dried, and then distilled under reduced pressure. The residue was purified by silica column chromatography, to obtain Compound 12 7.7 mg (70%). MS m/z 431 [M+H]+
The finely broken molecular sieve (100 mg) was mixed in anhydrous dichloromethane (5 ml), and then cooled to −20° C. (−)-Diethyl-tartrate (1.5 mg, 0.2 eq) and Ti(OiPr)4 (2.1 mg, 0.2 eq) were added thereto, followed by stirring for 30 minutes. Phorbaketal A (15 mg, 0.037 mmol) was put therein, followed by further stirring for 30 minutes. Tert-butyl hydroperoxide (TBHP, 4 mg, 1.2 eq) was put therein, followed by stirring for 3 hours, and then the progress of the reaction was confirmed by TLC. The reaction was terminated by using water (10 ml) at 0° C., and the stirring was further conducted at 0° C. for 1 hour. 30% of an aqueous NaOH solution (2 ml) and an aqueous NaCl solution (2 ml) were mixed therewith, followed by further stirring for 30 minutes, and then the cellite layer was filtered. The cellite layer was washed with dichloromethane, and the organic solvent was distilled under reduced pressure, and then the obtained residue was separated by silica column chromatography to obtain Compound 13. MS m/z 415 [M+H]+
Compound 11 (10 mg, 0.024 mmol) was dissolved in THF (5 ml), and then diisopropylamine (1.2 mmol) was mixed therewith, followed by stirring for 4 hours. The temperature of the reaction liquid was lowered to 0° C., and the reaction was terminated by water (20 ml). The organic solvent layer was separated, dried over anhydrous Na2SO4, and then distilled under reduced pressure, and the obtained residue was purified by silica column chromatography, to obtain Compound 14. MS m/z 516 [M+H]+
Phorbaketal A (10 mg, 0.025 mmol) and imidazole (3.4 mg, 0.05 mmol) were dissolved in DMF (10 ml), and then tert-butyldiphenyl silylchloride (8.2 ml, 0.03 mmol) was added at 0° C., followed by stirring for 6 hours. After the reaction was finished, the reaction liquid was layer-separated by using water and ethylacetate (2*15 ml), and the organic solvent layer was dried over Na2SO4. After distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 15. MS m/z 637 [M+H]+
Compound 13 (10 mg, 0.015 mmol) was dissolved in dichloromethane (1 ml), and DIBAL-H ((1-Bu2AlH)2, 40 ul, 1.5M in toluene) was added at −78° C., followed by further stirring for 30 minutes. The reaction was terminated by using ethanol (10 μl), and water (10 ml) and NaF (20 mg) were added thereto. The organic solvent layer was separated, dried, and distilled under reduced pressure, and the obtained residue was purified by silica column chromatograph, to obtain Compound 16. MS m/z 639 [M+H]+
Compound 17 was obtained by using Compound 16 (10 mg, 0.016 mmol) as a start material through the same synthesis method as Compound 8. MS m/z 663 [M+H]+
Compound 18 was obtained by using Compound 17 (10 mg, 0.015 mmol) as a start material through the same synthesis method as Compound 9. MS m/z 796 [M+H]+
Tetra-n-butylammonium fluoride (TBAF, CH3CH2CH2CH2)4N+F−) (4 mg, 0.015 mmol) was added in Compound 18 (10 mg, 0.013 mmol) dissolved in anhydrous THF (10 ml), followed by stirring at room temperature for 4 hours. The reaction liquid was layer-separated by using an aqueous saturated ammonium chloride solution and ethylacetate, and the obtained organic solvent layer was dried over Na2SO4. After distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 19. MS m/z 558 [M+H]+
Compound 20 was obtained by using Compound 15 (10 mg, 0.016 mmol) through the same synthesis method as Compound 1. MS m/z 793 [M+H]+
Compound 21 was obtained by using Compound 20 (10 mg, 0.013 mmol) through the same synthesis method as Compound 2. MS m/z 664 [M+H]+
Compound 22 was obtained by using Compound 21 (10 mg, 0.015 mmol) as a start material through the same synthesis method as Compound 10. MS m/z 766 [M+H]+
Compound 23 was obtained by using Compound 22 (10 mg, 0.013 mmol) as a start material through the same synthesis method as Compound 19. MS m/z 528 [M+H]+
Compound 4 (10 mg, 0.025 mmol) was dissolved in pyridine (5 ml), and then ethylcyanoacetate (5.7 mg, 2 eq) and a small amount of piperidine were added. The reaction liquid was stirred for 8 hours, and the progress of the reaction was confirmed by using TLC. The reaction was terminated by using water and dichloromethane, followed by layer separation. The organic solvent layer was distilled under reduced pressure, and the obtained residue was purified by silica column chromatography, to obtain Compound 24. MS m/z 492 [M+H]+
Compound 4 (10 mg, 0.025 mmol) and o-phenylene diamine (2.3 mg, 0.025 mmol) were dissolved in a water/acetonitrile (1:1) mixture liquid (3 ml), and then clayzic (20 mg) was put therein, followed by sufficient stirring at room temperature. Extraction with dichloromethane, washing with water, drying over Na2SO4, and then distillation under reduced pressure were conducted. The obtained residue was purified by silica column chromatography, to obtain Compound 25. MS m/z 485 [M+H]+
Compound 26 (9.4 mg, 69%) was obtained by using Compound 4 (10 mg, 0.025 mmol) as a start material through the same synthesis method as Compound 25. MS m/z 543 [M+H]+
Compound 26 (10 mg, 0.018 mmol) and NaOH (1.5 mg, 0.036 mmol) were dissolved in methanol, followed by stirring at 50° C. for 5 hours. After the reaction was terminated, neutralization was conducted by using water and hydrochloric acid, and then layer-separation was conducted by using ethylacetate (25 ml). The organic solvent layer was dried over anhydrous Na2SO4 and distilled under reduced pressure. The obtained residue was purified by silica column chromatography, to obtain compound 27 (6.16 mg, 65%). MS m/z 529 [M+H]+
Compound 4 (10 mg, 0.025 mmol) was dissolved in ethanol and water (7:3) solution (3 ml), and then NH2OH.HCl (3.45 mg, 2 eq) and NaHCO3 (4.2 mg, 2 eq) were added, followed by stirring at room temperature for 2 hours. Ice water (30 ml) was poured into the reaction liquid, and then layer-separation was conducted by using ethylacetate (2*25 ml). The organic solvent layer was dried over anhydrous Na2SO4 and distilled under reduced pressure. The obtained residue was purified by silica column chromatography, to obtain Compound 28 (8.83 mg, 86%). MS m/z 412 [M+H]+
NaOH (0.7 mg, 0.03 mmol) was put in diethylether (5 ml), and then nitrogen conditions were made. Triethyl phosphono acetate (6.7 mg, 0.03 mmol) dissolved in diethylether (2 ml) was put therein, and the mixture was stirred for 20 minutes and then further stirred for 30 minutes at 0° C. Compound 4 (10 mg, 0.025 mmol) was dissolved in ether (20 ml), and the reaction liquid was slowly added thereto, followed by stirring for 4 hours. Water (20 ml), hydrochloric acid (5 ml), and ether (4*20 ml) were added thereto. The obtained organic solvent layer was dried over MgSO4, and then distilled under reduced pressure. The obtained residue was purified by silica column chromatography, to obtain Compound 29 (9.1 mg, 78%). MS m/z 467 [M+H]+
Compound 30 (6.9 mg, 74%) was obtained by using Compound 29 (10 mg, 0.021 mmol) as a start material through the same synthesis method as Compound 27. MS m/z 439 [M+H]+
Compound 31 was obtained by using Compound 30 (10 mg, 0.022 mmol) as a start material through the same synthesis method as Compound 6. MS m/z 506 [M+H]+
A mixture of Compound 28 (10 mg, 0.024 mmol) and N-chlorosuccinimide (3 mg, 0.024 mmol) was dissolved in dichloromethane (3 ml), and then stirred at room temperature for 5 minutes. After further stirring for 30 minutes, layer-separation was conducted by using water (20 ml) and dichloromethane (2*20 ml). The organic solvent layer was dried over anhydrous Na2SO4. After distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 32. MS m/z 446 [M+H]+
A mixture of Compound 28 (10 mg, 0.024 mmol) and N-chlorosuccinimide (3 mg, 0.024 mmol) was dissolved in dichloromethane (3 ml), and then stirred at room temperature for 5 minutes. After Compound 32 was confirmed by TLC, phenylacetylene (10 mg, 0.1 mmol) and triethylamine (0.028 mmol) were added to the reaction liquid. After further stirring for 30 minutes, layer-separation was conducted by using water (20 ml) and dichloromethane (2*20 ml). The organic solvent layer was dried over anhydrous Na2SO4. After distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 33. MS m/z 512 [M+H]+
Phorbaketal acetate (10 mg, 0.02 mmol) dissolved in methanol (4 ml) solvent was allowed to react by using 10% Pd/C (5 mg) under hydrogen conditions for 10 hours. After filtering using cellite and washing with methanol, the obtained organic solvent layer was distilled under reduced pressure. The obtained residue was purified by silica column chromatography, to obtain Compound 34 (9 mg, 91%). MS m/z 451 [M+H]+
Hydrogen peroxide (4.6 μl, 0.068 mmol) and phorbaketal acetate (10 mg, 0.02 mmol) were allowed to react with methanol (10 ml) and 6N sodium peroxide (7.5 μl, 0.045 mmol) at 0° C. for 6 hours. The layer-separation was conducted by using dichloromethane and water, and the organic solvent layer was dried over Na2SO4. After distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 35 (8 mg, 87%). 1H-NMR (300 MHz, CDCl3): 1.27 (s, 3H), 1.49 (s, 3H), 1.62 (s, 3H), 1.98-1.86 (m, 8H), 2.08-2.88 (m, 5H), 2.54-2.69 (m, 2H), 3.48 (s, 1H), 4.09 (br s, 2H), 4.75 (br s, 2H), 5.11 (br s, 1H), 5.26 (s, 1H), 5.29 (br s, 1H), 5.58 (br s, 1H). MS m/z 415 [M+H]+
NaBH4 (1 mg, 0.033 mmol) was dissolved in methanol (6 ml), which was then mixed with Compound 35 (7 mg, 0.016 mmol) at 0° C. After the reaction liquid was stirred at room temperature for 2 hours, the solvent was distilled under reduced pressure, and the remaining residue was extracted with dichloromethane. After drying over anhydrous Na2SO4 and distillation under reduced pressure, the residue was purified by silica column chromatography, to obtain Compound 36 (6 mg, 92%). MS m/z 417 [M+H]+
Compound 36 (5 mg, 0.012 mmol), TPP (12 mg, 0.048 mmol), and N-chlorosuccineimide (6 mg, 0.48 mmol) were mixed with toluene solvent, and then allowed to react at 80° C. for 6 hours. After the reaction was terminated, the solvent was distilled under reduced pressure, and the residue was purified by silica column chromatography to obtain Compound 37 (5 mg, 93%). MS m/z 453 [M+H]+
Phorbaketal acetate (10 mg, 0.02 mmol), together with benzenthiol (9.3 μl, 0.09 mmol) and triethylamine (0.05 ml), was put in THF solvent, and then heated for 10 hours. After the reaction was terminated, the reaction liquid was layer-separated by using water (20 ml) and ethylacetate. The organic solvent layer was dried, followed by purification by silica column chromatograph, to obtain Compound 38. MS m/z 551 [M+H]+
DIABL (0.5 ml) was added to phorbaketal acetate (30 mg, 0.06 mmol) dissolved in anhydrous THF at −78° C. A saturated ammonium chloride solution was put therein, and then the layer-separation was conducted by using ethylacetate (2*20 ml). The obtained organic solvent layer was dried by sodium sulfate, followed by distillation under reduced pressure. The obtained residue was used to advance a subsequent reaction. The obtained residue (17 mg) was slowly mixed with anhydrous dichloromethane (10 ml) and triethylamine (0.2 ml) at 0° C. The reaction, together with mesyl chloride, was allowed to proceed at room temperature for 1 hour, and then the layer-separation was conducted by using dichloromethane and water, followed by concentration of the organic solvent layer. The residue was purified by using silica column chromatography, to obtain a product 18 mg. This product (10 mg) was reacted with N-dimethylaminopropylamine (81 mg, 0.78 mmol), together with Cs2CO3 (100 mg, 0.3 mmol), in DMF at 70° C. for 4 hours. Column chromatography was used to obtain Compound 39. MS m/z 541 [M+H]+
Compound 37 (7 mg, 0.014 mmol) and Cs2CO3 (50 mg) were mixed with DMF (5 ml), and then, together with phenylpiperazine (100 mg, 0.61 mmol), stirred at 80° C. for 6 hours. The solvent was removed by distillation under reduced pressure, and then the residue was purified by silica column chromatography, to obtain Compound 40. MS m/z 542 [M+H]+
NaBH4 (1 mg, 0.033 mmol) was slowly added into a methanol (6 ml) of cerium chloride heptahydrate (12 mg, 0.033 mmol) and phorbaketal acetate (10 mg, 0.02 mmol). After the reaction liquid was stirred at room temperature for 4 hours, the solvent was distilled under reduced pressure. The residue was layer-separated by using dichloromethane and water, and then the organic solvent layer was dried over Na2SO4, followed by proceeding to the next step. The residue was dissolved in dichloromethane, and then a 85% tetrafluorboric acid diethylether complex and phenylpiperazine were slowly added at −60° C. while stirring for 1 hour. The mixture was layer-separated by using water and dichloromethane, followed by distillation under reduced pressure, and the organic solvent layer was separated and purified by using silica column chromatography, to obtain Compound 41. MS m/z 587 [M+H]+
Compound 41 (3 mg, 0.005 mmol) was dissolved in methanol (2 ml), and then K2CO3 (1 mg) was added thereto. The reaction was stirred at room temperature for 1 hour, followed by filtration and distillation under reduced pressure, to obtain a residue. The residue was purified by using silica column chromatography, to obtain Compound 42. MS m/z 545 [M+H]+
Phorbaketal acetate (10 mg, 0.02 mmol) was dissolved in anhydrous THF, and then
(1 ml) was added at −30° C., followed by stirring. The layer-separation was conducted by using an aqueous saturated ammonium chloride solution and ethylacetate (2*20 ml), and the separated organic solvent layer was dried over anhydrous Na2SO4. After distillation under reduced pressure, the residue was purified by silica column chromatography, to obtain Compound 43 (8 mg, 82%). 1H-NMR (300 MHz, CDCl3): d1.52 (s, 3H), 1.56-1.86 (m, 9H), 2.08-2.88 (m, 8H), 2.54-2.69 (m, 2H), 4.3 (br s, 2H), 4.75-4.83 (m, 2H), 5.11-5.21 (m, 3H), 5.24-5.27 (m, 2H), 5.64-5.72 (m, 3H), 5.82-5.95 (m, 2H). MS m/z 427 [M+H]+
Trimethylammonium (37 μl, 0.07 mmol, 2M solution) was mixed with dichloromethane (6 ml), which was then cooled to 0° C. Benzenethiol (79 μl, 0.077 mmol) was slowly added, and then the mixture liquid was stirred for 20 minutes. Compound 4 (10 mg, 0.02 mmol) was slowly added at −78° C., followed by further stirring for 15 minutes. THF (4 ml) was added, followed by stirring for 5 minutes, and then acetaldehyde (43 μl, 0.077 mmol) was slowly added. After further stirring for 20 minutes, the layer separation was carried out by adding water (5 ml) and dichloromethane (5 ml) thereto. The organic solvent layer was washed with 1N HCl (5 ml), and further, the aqueous layer was extracted by using ethylacetate (2*5 ml). The entire organic solvent layer was collected, washed with water (10 ml) and salt water (10 ml), and then dried over MgSO4. After distillation under reduced pressure, the obtained residue was separated by using column chromatography to obtain Compound 44. MS m/z 459 [M+H]+
Starting from phorbaketal A (20 mg, 0.05 mmol) as a start material, acetic acid was allowed to react in the cyclohexane solvent using sulfuric acid as a catalyst, to thereby transform free alcohol into an acetyl group. The obtained product was used by using the same synthesis method as Compound 12 to obtain Compound 45 (19 mg, 95%). MS m/z 443 [M+H]+
Triethylamine (1 ml) and Compound 45 (15 mg, 0.037 mmol) were dissolved in anhydrous dichloromethane (10 ml), and then methanesulfonyl chloride (8.6 μl, 0.11 mmol) was added thereto, followed by stirring at room temperature for 1 hour at 0° C. The reaction liquid was layer-separated by using water (20 ml) and dichloromethane (50 ml), and the organic solvent layer was washed with water and salt water, followed by drying over Na2SO4. Distillation under reduced pressure and separation by silica column chromatography were conducted to obtain Compound 46 (20 mg, 94%). 1H-NMR (300 MHz, CDCl3): d1.56 (s, 3H) 1.62 (s, 3H) 1.69 (s, 3H) 1.73 (s, 3H), 1.78 (s, 3H), 1.90 (s, 3H), 1.98-2.05 (m, 11H), 2.46 (brd, 1H), 4.44 (brs, 2H), 4.55-4.70 (d, 2H), 4.76-4.80 (m, 1H), 5.07-5.11 (m, 1H), 5.25 (s, 1H), 5.29 (s, 1H), 5.68 (brs, 1H), 5.69 (brs, 1H). MS m/z 521 [M+H]+
Compound 46 (15 mg, 0.026 mmol) was dissolved in THF (10 ml), and then the temperature was lowered to 0° C. Histamine (59 mg, 0.53 mmol) was added, followed by stirring at 80° C. for 7 hours. The layer separation was carried out by using ethylacetate (50 ml) and water (3*10 ml), followed by drying over Na2SO4. After distillation under reduced pressure, the obtained residue was purified by silica column chromatography, to obtain Compound 47. MS m/z 536 [M+H]+
Phorbaketal A (5 mg, 0.01 mmol) and triethylamine (1 ml) were dissolved in dichloromethane, and then 2-chloro-2,2-diphenylacetylchloride (100 mg, 0.37 mmol) was added, followed by stirring at room temperature for 10 hours. After the reaction was terminated, the layer-separation was conducted by using dichloromethane and water. The organic solvent layer was distilled under reduced pressure, and then the obtained residue was purified by silica column chromatography, to obtain Compound 48. MS m/z 627 [M+H]+
Phorbaketal A (10 mg, 0.025 mmol) and triethylamine (1 ml) were mixed with anhydrous dichloromethane (10 ml) at 0° C., and then 2-carbomethoxy-3-thiophene sulfonyl chloride (100 mg, 0.41 mmol) was added, followed by stirring at room temperature for 12 hours. The layer separation was carried out by using water (20 ml) and dichloromethane (50 ml), and the organic solvent layer was distilled under reduced pressure. The obtained residue was purified by silica column chromatography, to obtain Compound 49 (13 mg, 89%) MS m/z 603 [M+H]+
Compound 49 (10 mg, 0.016 mmol) was mixed with lithium hydroxide (1N, 10 μl) dissolved in dioxane (10 ml), followed by stirring at room temperature for 1 hour. After oxidation using an aqueous 2N HCl solution, separation by silica column chromatography was conducted to obtain Compound 50. MS m/z 589 [M+H]+
Phorbaketal A (10 mg, 0.025 mmol) and triethylamine (1 ml) were mixed with anhydrous dichloromethane (10 ml) at −20° C., and then triflic anhydride (0.015 ml, 0.088 mmol) was added, followed by stirring for 30 minutes. The reaction liquid was diluted with dichloromethane (20 ml), and washed with a cooled aqueous 1N HCl solution, NaHCO3, salt water, and water. The organic solvent layer was separated, dried over Na2SO4 and distilled under reduced pressure. The residue was mixed with DIPEA (1 ml) and THF (10 ml), and then thiomorpholine (200 mg) was added thereto at 0° C. After stirring for 4 hours, the layer separation was carried out by using ethylacetate (100 ml) and water (3*10 ml). The organic solvent layer was dried over Na2SO4, and then purified by silica column chromatography, to obtain Compound 51 (8 mg, 71%). MS m/z 484 [M+H]+
Phorbaketal A (20 mg, 0.050 mmol), together with triethylamine (1 ml), was put in anhydrous dichloromethane (10 ml) at 0° C., which was then allowed to react with diphenyl acetyl chloride (100 mg, 0.43 mmol), followed by stirring for 10 hours. The layer-separation was conducted by using water (20 ml) and dichloromethane (30 ml), followed by washing with salt water and water, drying over Na2SO4, distillation under reduced pressure, and separation by silica column chromatography, to obtain Compound 52. MS m/z 593 [M+H]+
NaBH4 (1 mg, 0.033 mmol) was mixed with phorbaketal A (10 mg, 0.025 mmol) and methanol (20 ml) at 0° C. The reaction liquid was stirred for 4 hours, and then the solvent was distilled under reduced pressure. The residue was extracted with dichloromethane (3*20 ml), followed by drying over Na2SO4 and then distillation under reduced pressure. The residue was purified by using silica column chromatography, to obtain Compound 53. MS m/z 403 [M+H]+
Phorbaketal A (12 mg, 0.03 mmol) and carbon tetrabromide (20 mg, 0.36 mmol), together with pyridine (0.5 ml), were mixed with triethylphosphate (12 μl, 0.075 mmol) in anhydrous dichloromethane (5 ml) at 0° C., and the mixture was allowed to react at room temperature for 9 hours. The reaction liquid was washed with 10% HCl and an aqueous saturated NaHCO3 solution, and then the layer separation was carried out by using water and dichloromethane. The organic solvent layer was dried over Na2SO4. Purification using column chromatography was conducted to obtain Compound 54 (10 mg, 63%). MS m/z 534 [M+H]+
Compound 54 (9 mg, 0.016 mmol), together with 2,4,6-colidine (25 μl) and bromotrimethylsilane (25 μl, 0.16 mmol), was allowed to react with anhydrous dichloromethane (3 ml) at 0° C. After the reaction was allowed to proceed at room temperature for 15 hours, the solvent was removed, and then the residue was allowed to react with 2N NaOH (0.8 ml) at room temperature for 4 hours. The solvent was removed, and column chromatography was used to obtain Compound 55 (7 mg, 74%). MS m/z 555 [M+H]+
Phorbaketal A (20 mg, 0.05 mmol) and THF SO3 pyridine complex (20 mg, 0.13 mmol) were slowly mixed with each other at 0° C. After stirring at room temperature for 1 hour, the reaction liquid was purified by silica gel column chromatography, to obtain Compound 56. MS m/z 517 [M+H]+
Phorbaketal A (15 mg, 0.037 mmol) and triethylamine (1 ml) were mixed with anhydrous dichloromethane (10 ml) at 0° C., and then methane sulfonyl chloride (8.6 μl, 0.11 mmol) was added, followed by stirring at room temperature for 1 hours. The layer separation was carried out by using water (20 ml) and dichloromethane (20 ml), and the organic solvent layer was distilled under reduced pressure. The obtained residue was purified by silica column chromatography, to obtain Compound 57. MS m/z 477 [M+H]+
Phorbaketal A (10 mg, 0.025 mmol) and Hg(OAC)3 were put and allowed to react with each other in a mixture solvent of THF (5 ml) and water (1 ml) for 2 hours. After THF was removed by distillation under reduced pressure, water (5 ml) was added and NaOH (20 mg) and NaBH4 (910 mg) were added, followed by reaction for 30 minutes. The layer separation was carried out by using water (10 ml) and dichloromethane (20 ml), and the organic solvent layer was distilled under reduced pressure. The obtained residue was purified by silica column chromatography, to obtain Compound 58. MS m/z 417 [M+H]+
Phorbaketal A (15 mg, 0.038 mmol), 2-thiophene carbonylchloride (5.50 mg, 1 eq), and DIEA(N,N-diisopropylethylamine) (0.01 mmol) were mixed with dichloromethane, followed by stirring at room temperature for 2 hours. Ice water (30 ml) was poured into the reaction liquid, and then reaction was terminated by using ethylacetate (2*25 ml). The organic solvent layer was collected, and then dried by using anhydrous sodium sulfate and distilled under reduced pressure. The obtained residue was purified by silica column chromatography to obtain Compound 59 (16.5 mg, 86%). 1H-NMR (300 MHz, CDCl3): d 1.60 (3H, s) 1.68 (3H, s) 1.76 (3H, s) 1.78 (3H, s) 1.84 (3H, s) 2.07-2.20 (6H, m) 2.56-2.74 (3H, t) 4.64 (1H, s), 4.70-4.88 (3H, t) 5.10 (1H, s) 5.26 (1H, d) 5.36 (1H, s), 5.77 (1H, s) 6.66 (1H, s) 7.12 (1H, m) 7.57 (1H, d) 7.81 (1H, s). MS m/z 509 [M+H]+
NaH (0.72 mg, 1.2 eq), phorbaketal A (10 mg, 0.025 mmol), and 2,2-difluorobenzylbromide (5.2 mg, 1 eq) were put in THF (5 ml), followed by stirring at 0° C. for 4 hours. The reaction was terminated by using water and ethylacetate (3*25 ml), and the obtained organic solvent layer was dried by using sodium sulfate. After distillation under reduced pressure, the residue was purified by silica column chromatography, to obtain Compound 60 (11.1 mg, 85%). H-NMR (300 MHz, CDCl3): d1.60 (3H, s) 1.68 (3H, s) 1.74 (3H, s) 1.76 (3H, s) 1.82 (3H, s) 2.04-2.12 (6H, m) 2.47-2.63 (2H, t) 3.92-4.10 (2H, m), 4.55 (2H, s) 4.76 (3H, t) 5.09 (1H, s) 5.26 (1H, d) 5.32 (1H, s), 5.66 (1H, s) 6.64 (1H, s) 6.88 (2H, m) 7.22 (1H, m). MS m/z 525 [M+H]+
Compound 61 was obtained at a yield of 91% by employing the same synthesis method as Compound 60 while 2,3,5,6-tetrafluoromethyl-4-trifluoromethane-benzylbromide was used in stead of 2,2-difluorobenzylbromide. 1H-NMR (300 MHz, CDCl3): d1.61 (3H, s) 1.68 (3H, s) 1.75 (3H, s) 1.76 (3H, s) 1.82 (3H, s) 2.08-2.11 (6H, m) 2.51-2.76 (3H, m) 4.47 (2H, s) 4.53-4.64 (3H, m), 4.80 (1H, s) 5.10 (1H, s) 5.27 (1H, d) 5.35 (1H, s), 5.78 (1H, s) 6.68 (1H, d). MS m/z 629 [M+H]+
Compound 5 (10 mg, 0.025 mmol) was dissolved in an ethanol/water (7:3) mixture liquid (3 ml), and then NH2OHHCl (3.9 mg, 2.2 eq) and NaOH (4.0 mg, 4 eq) were put therein. Then, the reaction was allowed to proceed at room temperature for 2 hours. The reaction liquid was put in ice water (30 ml) and the layer separation was carried out by using ethylacetate (2*25 ml). The organic solvent layer was collected, and then dried by using NaSO4. The organic solvent was distilled under reduced pressure, and the obtained residue was purified by column chromatography to obtain Compound 62. MS m/z 427 [M+H]+
Phorbaketal acetate (10 mg, 0.022 mmol) was dissolved in an ethanol/water (7:3) mixture liquid (3 ml), and then NH2OHHCl (1.5 mg, 1 eq) and NaOH (1.8 mg, 2 eq) were put therein. Then, the reaction was allowed to proceed at room temperature for 2 hours. The reaction liquid was put in ice water (30 ml) and the layer separation was carried out by using ethylacetate (2*25 ml). The organic solvent layer was collected, and then dried by using Na2SO4. The organic solvent was distilled under reduced pressure, and the obtained residue was purified by column chromatography to obtain Compound 63. MS m/z 414 [M+H]+
Phorbaketal acetate (10 mg, 0.022 mmol) was put in CH2Cl2 (5 ml) at 0° C., and m-CPBA (meta-chloroperoxybenzoic acid) (3.9 mg, 1 eq) was dissolved in CH2Cl2 (6 ml), which were then slowly mixed. After the reaction at room temperature for 3 hours, an aqueous saturated NaHCO3 solution was added thereto, followed by stirring for 30 minutes. The layer separation was carried out by using CH2Cl2, and then the organic solvent layer was dried. After distillation under reduced pressure, the residue was separated by column chromatography, to obtain Compound 64.
Compound 38 (6 mg) was dissolved in methanol (2 ml), which was then mixed with K2CO3 (1 mg), followed by stirring for 1 hour. The reaction liquid was subjected to filtration, concentration, and silica column chromatography, to obtain Compound 65. 1H-NMR (300 MHz, CDCl3)d: 1.27 (d, 3H), 1.50 (s, 3H), 1.58 (s, 3H), 1.65 (s, 3H), 1.72 (s, 3H), 1.80 (s, 3H), 1.98-2.05 (m, 2H), 2.11 (s, 6H), 2.61 (dd, 1H), 2.94 (br ddd, 1H), 3.42-3.48 (m, 1H), 3.78-3.81 (m, 1H), 4.34 (br s, 2H), 4.71-4.81 (m, 1H), 5.03-5.12 (m, 1H), 5.27 (d, 1H), 5.52 (s, 1H), 5.70 (br s, 1H) 7.22-7.31 (m, 5H). MS m/z 509 [M+H]+
Sodium hydride (0.7 mg, 1.2 eq) was added to a THF (5 ml) mixture liquid of phorbaketal A (10 mg, 0.025 mmol) and thiazole derivative
(7.72 mg, 1 eq) at 0° C., followed by stirring at room temperature for 4 hours. The layer separation was carried out by using water and ethylacetate (3*20 ml), and the organic solvent layer was separated, and then dried over anhydrous Na2SO4. The solvent was removed by distillation under reduced pressure, and the residue was purified by chromatography to obtain Compound 66. MS m/z 672 [M+H]+
Phorbaketal acetate (20 mg, 0.045 mmol) was dissolved in THF (4 ml), and then mixed with L-selectride (1.0M in THF, 0.07 ml) under nitrogen conditions while the temperature was lowered to −78° C. After the reaction liquid was stirred at −78° C. for 30 minutes, the temperature was raised to room temperature. An aqueous 10% NaOH solution was put in the reaction liquid, and then the organic solvent layer was separated. The aqueous layer was layer-separated by using ethylacetate (3 ml*5), and the separated organic solvent layer was collected and then dried over anhydrous Na2SO4. After distillation under reduced pressure, the residual was purified by column chromatography, to obtain Compound 67. MS m/z 401 [M+H]+
Copper cyanide (50 mg, 0.56 mmol) was dissolved in diethyl ether (10 ml), and then mixed with MeLi (1.6M in diethyl ether, 23 mg, 1.12 mol) at −10° C., followed by stirring for 1 hour. Compound 43 (15 mg, 0.033 mmol) wad added thereto, followed by stirring for 3 hours. After the layer separation was carried out by using an aqueous saturated ammonium chloride solution and ethylacetate, the organic solvent layer was dried, followed by separation by silica column chromatography, to obtain Compound 68. MS m/z 415 [M+H]+
Compound 37 (6 mg, 0.012 mmol) and Cs2CO3 (50 mg) were mixed with DMF (5 ml), and then, together with phenylpiperazine (1 ml, 12.19 mmol), stirred at 80° C. for 6 hours. The solvent was removed by distillation under reduced pressure, and then the residue was purified by silica column chromatography, to obtain Compound 69 (3 mg, 65%). MS m/z 452 [M+H]+
Compound 37 (6 mg, 0.012 mmol) and Cs2CO3 (50 mg) were mixed in isopropylamine (5 ml, 12.19 mmol), and the mixture was stirred for 6 hours while being heated. The solvent was removed by distillation under reduced pressure, and then the residue was purified by silica column chromatography, to obtain Compound 70. MS m/z 440 [M+H]+
Phorbaketal (20 mg, 0.05 mmol), α-D-glucopyranocyl bromide tetrabenzoate (82 mg, 0.125 mmol), and molecular sieve (0.5 g) were mixed with dichloromethane, and then silver triplet (10 mg, 0.03 mmol) was slowly mixed therewith at 0° C. The mixture was stirred at room temperature for 8 hours, followed by filtration, and washing with water, and distillation under reduced pressure. The obtained residue was purified by silica column chromatography to obtain Compound 71. MS m/z 561 [M+H]+
Phorbaketal (10 mg, 0.025 mmol), DCC(N,N′-dicyclohexylcarbodiimide) (6.18 mg, 0.030 mmol), and N-boc proline (0.62 mg, 0.025 mmol) were mixed with dichloromethane (5 ml) while 4-dimethylaminopyridine (DMAP) functioning as a catalyst was put therein, and then the stirring was conducted until an ester structure was completed at room temperature. The reaction liquid was washed to remove N,N-dicyclohexyl urea. The residue was washed with water (3*30 ml) and 5% acetic acid (3*30 ml) and again washed with water (3*30 ml), and then dried over anhydrous NaSO4. The solvent was removed, and the residue remaining after distillation under reduced pressure was purified by column chromatography, to obtain Compound 72. MS m/z 596 [M+H]+
The following compounds (Compounds 73˜81) were separated and purified from Phorbas sponge by the following procedure. The sponge was freeze-dried, and then the dry weight 500 g was subjected to extraction with a methanol and
CH2Cl2 (1:1) mixture solution. The extraction liquid was layer-separated by using water and CH2Cl2. The organic solvent layer was layer-separated by using an aqueous 90% methanol solution and n-hexane. The aqueous 90% methanol solution layer was fractionized by using reverse phase silica column chromatography. Hereinafter, specific separation conditions of the examples are as follows.
1H-NMR (300 MHz, CDCl3)d: 1.29 (s, 6H), 1.75 (s, 3H), 1.76 (s, 3H) 1.81 (s, 3H), 1.87 (m, 1H), 2.04 (m, 1H), 2.43 (m, 1H), 2.57 (s, 1H), 2.59 (s, 1H), 2.77 (s, 1H), 4.06 (s, 2H), 4.49 (m, 1H), 4.75 (s, 1H), 5.27 (d, 1H), 5.29 (s, 1H), 5.54 (s, 1H), 5.61 (d, 1H), 5.66 (d, 1H), 6.70 (m, 1H); MS m/z 431 [M+H]+
1H-NMR (300 MHz, CDCl3)d: 1.14 (s, 3H), 1.17 (s, 3H), 1.40 (d, 1H), 1.76 (s, 3H), 1.78 (m, 1H), 1.81 (s, 3H), 1.82 (s, 3H), 1.87 (m, 1H), 2.05 (m, 1H), 2.08 (m, 1H), 2.19 (m, 1H), 2.33 (m, 1H), 2.44 (m, 1H), 2.58 (m, 1H), 2.59 (m, 1H), 3.27 (m, 1H), 4.07 (s, 2H), 4.51 (m, 1H), 4.77 (m, 1H), 5.28 (d, 1H), 5.30 (s, 1H), 5.55 (s, 1H), 6.71 (m, 1H); MS m/z 433 [M+H]+
1H-NMR (300 MHz, CDCl3)d: 1.11 (s, 1H), 1.18 (s, 1H), 1.29 (s, 1H), 1.44 (m, 1H), 1.51 (s, 1H), 1.67 (s, 1H), 1.69 (s, 1H), 1.76 (s, 1H), 1.81 (s, 1H), 1.83 (m, 1H), 2.01 (s, 1H), 2.02 (s, 1H), 2.44 (m, 1H), 2.59 (m, 1H), 2.61 (s, 1H), 2.63 (s, 1H), 4.00 (s, 1H), 4.08 (m, 2H), 4.49 (m, 1H), 5.24 (s, 1H), 5.51 (s, 1H), 6.66 (m, 1H); MS m/z 415 [M+H]+
1H-NMR (300 MHz, CDCl3) d: 1.07 (s, 3H), 1.08 (s, 3H) 1.76 (s, 3H), 1.80 (s, 3H), 1.82 (s, 3H), 1.86 (m, 1H), 2.01 (m, 1H), 2.03 (s, 1H), 2.04 (m, 1H), 2.28 (m, 2H), 2.44 (m, 1H), 2.59 (m, 1H), 2.60 (s, 1H), 2.67 (m, 2H), 4.05 (s, 1H), 4.08 (s, 2H), 4.50 (m, 1H), 4.74 (m, 1H), 5.24 (m, 1H), 5.30 (s, 1H), 5.54 (s, 1H), 6.71 (m, 1H); MS m/z 415 [M+H]+
1H-NMR (300 MHz, CDCl3)d: 1.27 (s, 6H), 1.75 (s, 3H), 1.76 (s, 3H), 1.81 (s, 3H), 1.86 (m, 1H), 2.04 (m, 1H), 2.43 (m, 1H), 2.57 (m, 1H), 2.58 (m, 1H), 2.75 (d, 1H), 4.06 (br, 2H), 4.49 (m, 1H), 4.75 (m, 1H), 5.27 (d, 1H), 5.29 (s, 1H), 5.53 (s, 1H), 5.60 (m, 1H), 5.66 (m, 1H), 6.09 (m, 1H); MS m/z 415 [M+H]+
1H-NMR (300 MHz, CDCl3)d: 1.27 (s, 6H), 1.75 (s, 3H), 1.76 (s, 3H), 1.81 (s, 3H), 1.86 (m, 1H), 2.04 (m, 1H), 2.43 (m, 1H), 2.57 (m, 1H), 2.58 (m, 1H), 2.75 (d, 1H), 4.49 (m, 1H), 4.60 (m, 2H), 4.75 (m, 1H), 5.27 (d, 1H), 5.29 (s, 1H), 5.59 (s, 1H), 5.60 (m, 1H), 5.66 (m, 1H), 6.09 (m, 1H); MS m/z 457 [M+H]+
1H-NMR (300 MHz, CDCl3)d: 1.76 (s, 3H), 1.80 (s, 3H), 1.82 (s, 3H), 1.86 (s, 3H), 1.86 (m, 1H), 2.01 (m, 1H), 2.03 (s, 1H), 2.04 (m, 1H), 2.34 (t, 2H), 2.44 (m, 1H), 2.59 (m, 1H), 2.60 (s, 1H), 2.90 (s, 2H), 4.05 (s, 2H), 4.08 (s, 2H), 4.50 (m, 1H), 4.74 (m, 1H), 5.24 (m, 1H), 5.30 (s, 1H), 5.54 (s, 1H), 5.86 (d, 1H), 6.09 (s, 1H), 6.71 (m, 1H); MS m/z 413 [M+H]+
1H-NMR (300 MHz, CDCl3)d: 1.65 (m, 2H), 1.71 (s, 3H), 1.75 (s, 3H), 1.79 (s, 3H), 1.81 (s, 3H), 1.86 (m, 1H), 2.04 (m, 1H), 2.11 (m, 2H), 2.43 (m, 1H), 2.57 (m, 1H), 2.58 (m, 1H), 3.99 (m, 1H), 4.06 (br, 2H), 4.49 (m, 1H), 4.75 (m, 1H), 4.82 (s, 1H), 4.92 (s, 1H), 5.27 (d, 1H), 5.29 (s, 1H), 5.53 (s, 1H), 6.09 (m, 1H); MS m/z 415 [M+H]+
1H-NMR (300 MHz, CDCl3)d: 1.65 (m, 2H), 1.71 (s, 3H), 1.75 (s, 3H), 1.79 (s, 3H), 1.81 (s, 3H), 1.86 (m, 1H), 2.04 (m, 1H), 2.11 (m, 2H), 2.43 (m, 1H), 2.57 (m, 1H), 2.58 (m, 1H), 3.99 (m, 1H), 4.06 (br, 2H), 4.49 (m, 1H), 4.75 (m, 1H), 4.82 (s, 1H), 4.92 (s, 1H), 5.27 (d, 1H), 5.29 (s, 1H), 5.53 (s, 1H), 6.09 (m, 1H); MS m/z 415 [M+H]+
Animal cell line CV-1 was used in transfection search. Cells were cultured in DMEM medium within a cell incubator containing 5% carbon dioxide at 37° C. The medium contained 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. On day 1 of the experiment, CV1 cells were seeded in a 96-well plate at 5,000 cells/well. On day 2, the seeded cells were transfected with plasmid expressing GAL-mNurr1, plasmid expressing luciferase gene, and plasmid expressing β-galactosidase by using a transfection reagent, Superfect (QIAGEN). After 16 hours, the transfected cells were treated with compounds of Chemical Formula I dissolved in dimethylsulfoxide (DMSO) by the concentrations. Cells treated with dimethylsulfoxide having the final concentration of 1% were used as a negative control group. After culturing for 24 hours, the cells were lysed by using a lysis buffer, and luciferin was added thereto to measure the luciferase activity by a luminometer. After addition of an ONPG reagent, β-galactosidase activity was measured by an ELISA reader. The measured luciferase value was corrected by the activity value of β-galactosidase. The experiment results showed that CMDD-X (
Animal cell line CV-1 was used in transfection search. Cells were cultured in DMEM medium within a cell incubator containing 5% carbon dioxide at 37° C. The medium contained 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. On day 1 of the experiment, CV1 cells were seeded in a 96-well plate at 5,000 cells/well. On day 2, the seeded cells were transfected with plasmid expressing GAL-hLXRa or GAL-hLXRB, plasmid expressing luciferase gene, and plasmid expressing β-galactosidase, by using a transfection reagent, Superfect (QIAGEN). After 16 hours, the transfected cells were treated with compounds of Chemical Formula I dissolved in dimethylsulfoxide (DMSO) by the concentrations (here, the final concentration of the LXR agonist, T0901517, was 500 nM). Cells treated with dimethylsulfoxide having the final concentration of 1% were used as a negative control group. After culturing for 24 hours, the cells were lysed by using a lysis buffer, and luciferin was added thereto to measure the luciferase activity by a luminometer. After addition of an ONPG reagent, β-galactosidase activity was measured by an ELISA reader. The measured luciferase value was corrected by the activity value of β-galactosidase. The experiment results showed that CMDD-X had an IC50 value of 20 nM on LXR. In addition, Compound 1, Compound 4, Compound 7, and Compound 8 showed IC50 values of 21 nM, 5 nM, 11 nM, and 85 nM, respectively. Other compounds of Chemical Formula I also showed similar antagonistic activity values similar to that of CMDD-X. In addition, in order to measure selectivity on various nuclear receptors (RXR, PPAR, PXR, CAR, ER, LRH-1, GR, NGFI-B, ERR, Rev-erb, GCNF, TR, HNF4, COUP, EAR, VDR, AR, DAX-1, TLX, and VDR), antagonistic activity on various nuclear receptors were measured by employing the same method (only agonists for respective nuclear receptors were exchanged). CMDD-X never showed antagonistic activity on the other nuclear receptors, and thus exhibited excellent selectivity. Other compounds of Chemical Formula I never showed antagonistic activities on the other nuclear receptors, and thus exhibited excellent selectivity. Accordingly, it was verified that the compounds of Chemical Formula I are antagonist compounds selective to LXR.
In the present example, in order to verify efficacy to prevent and treat diabetes of the compound of Chemical Formula I (CMDD-X or Compound 7) according to the present invention, BKS.Cg-+Leprdb/+Leprdb (db/db) mice as an insulin-independent diabetes animal model and general mice (C57BL/6J) fed with high-fat feedstuff were used. After the disease animal model was orally administered with CMDD-X, the diabetes-related index between the administered group and the control group was measured to verify the efficacy as an agent for preventing and treating diabetes. BKS.Cg-+Leprdb/+Leprdb (db/db) mice (male, 7-week old) as type 2 diabetes animal model were purchased and used. The experiment animals were adapted to a housing environment for 1 week while pellet type general diet (lab-chow) was supplied, and then divided into 2 groups of 18 mice each by randomized block design to have similar blood sugar and body weight. For general mice fed with high-fat feedstuff as another animal model, 6-week old C57BL/6J mice was purchased, and then were adapted to a lap environment for 2 weeks. After that, high-fat diet containing 35% fat diet (weight ratio, Research Diets. Co. Ltd.) was supplied for 14 weeks. The efficacy to prevent and treat diabetes by CMDD-X was observed through oral administration. A mouse fed with only 0.5% carboxylmethyl cellulose, as a medicine delivery, was used as a negative control group. The medicine was orally administered to the db/db mice for 4 weeks, a total of 28 days, and to the C57BL/6J mice, which was fed with the high-fat diet, for the last 6 weeks, at a dose of 10 mg/kg/day. The experiment animals after administration were fasted for 15 hours before autopsy, and then the blood taken from the orbital venous treated with heparin. After that, the plasma was separated by centrifugation of 1,000×g at 4° C. for 15 minutes, and then refrigerated at −70° C. until analysis was conducted to measure the insulin and Adiponectin concentrations. The pancreatic tissue was fixed in 10% formaldehyde solution containing PBS. The other organ tissues were rapidly cooled in the liquid nitrogen immediately after removal thereof, and kept at −70° C. and then analyzed. T-test was conducted with respect to the control group and the administered group, a significant difference of less than 5% (P<0.05) was determined as having have a statistical significance.
In order to measure the blood sugar reduction efficacy of the compound of Chemical Formula I (CMDD-X or Compound 7), the blood was collected through tail veins of the db/db mice before experiment ending, after feeding, and after stomach was empty for 15 hours, and the blood sugar was measured by using a glucose oxidase method. After food intake, the blood sugar of the CMDD-X-administered group was lower than that of the control group by 23.6%, and the blood sugar at the time of an empty stomach was reduced by about 26.3% (
The insulin and adiponectin concentrations of the plasma, obtained from the orbital venous of the db/db mice orally administered with CMDD-X (or Compound 7) for 4 weeks, were measured by using an Elisa Kit. The plasma insulin concentrations of the control group and the CMDD-X-administered group were shown in
On the end day of the experiment, the db/db mice were fasted for 15 hours, and then intraperitoneally administered with a glucose solution of 1 g per 1 kg body weight. The blood was collected from the tails of the db/db mice according to the time. The results of measurement of blood sugar by the times of the CMDD-X-administered group and the control group were shown in
On the end day of experiment, the disease animal with diabetes induced by high-fat diet were also fasted for 15 hours in the same manner as the db/db mice, and then intraperitoneally administered with a glucose solution of 1 g per 1 kg body weight. The blood was collected from the tails of the mice according to the time. The results of checking of the blood sugar level by the times of the CMDD-X-administered group and the control group were shown in
Gene expression analysis was conducted to establish the mechanism. The gene analysis was conducted by using micro-array and the Q-RT PCR method. In the mice fed with CMDD-X (or Compound 7), expression of Gyk as a glycolysis biomarker was increased (1.3 times), and expressions of LXR, SREBP1c, and SCD1, which are genes of synthesizing fatty acid by using decomposed glucose, were very increased (
In order to verify the efficacy to treat renal failure as a diabetes complication, a db/db disease animal model was used. 8-week old db/db mice were orally administered with the compound of Chemical Formula I (CMDD-X or Compound 7) at a dose of 10 mg/kg/day for 4 weeks, and then the blood urea nitrogen as a marker to treat renal failure was measured. The results of the CMDD-X-administered group were shown in
In order to verify efficacy to prevent and treat fatty liver of the compounds according to the present invention, a fatty liver occurrence model by a medicine, a fatty liver occurrence model by high fat, and a fatty liver occurrence model by methione-deficient diet (MCD) were used. The mice fed with general diet (chow diet) was simultaneously administered with a nuclear receptor LXR agonist (T0901317) to induce fatty liver, and then the efficacy to prevent and treat fatty liver by the compound of Chemical Formula I was tested. After the disease animal model was orally administered with CMDD-X (or Compound 7), the fatty liver-related index between the administered group and the control group was measured to verify the efficacy to prevent and treat fatty liver. The mouse fed with only 0.75% carboxylmethyl cellulose, as a medicine delivery, was used as a negative control group. First, the C57BL/6J mouse fed with general feedstuff was orally administered with the LXR agonist [T0901317 (10 mg/kg)] once per day for 5 days, to thereby induce fatty liver. In addition, the compound of Chemical Formula I (CMDD-X and Compound 7, 10 mg/kg) and the LXR agonist were simultaneously administered, to verify efficacy to prevent and treat fatty liver. The C57BL/6J mouse was fed with 35% high-fat feedstuff for 8 weeks, to induce fatty liver, and then CMDD-X (or Compound 7), together with high-fat diet, was orally administered at a dose of 10 mg/kg/day for 6 weeks. The ob/ob animal model was fed with MCD diet for 4 weeks to induce fatty liver and, at the same time, was orally administered with the compound of Chemical Formula I (CMDD-X or Compound 7, 10 mg/kg), to verify the efficacy to prevent and treat the occurrence of fatty liver. As an analysis result, fat absorption into the liver was inhibited in all of the fatty liver occurrence model by medicine, the fatty liver occurrence model by high fat, and the fatty liver occurrence model by MCD. Moreover, the blood LDL was decreased and the liver function was significantly improved by inhibiting fatty acid biosynthesis and decomposing the accumulated fatty acid to release heat. This resulted in the reduction in ALT, which is a liver-function biomarker, and the reduction in MCP-1 and TNFα, which are inflammatory cytokines (
In order to test the antiatherogenic efficacy of the compound of Chemical Formula I (e.g., CMDD-X or Compound 7) according to the present invention, Raw264.7 macrophage activated by LPS and HUVEC endotheliocyte activated by TNF-α were used. Raw264.7 macrophage was cultured in DMEM medium within a cell incubator containing 5% carbon dioxide at 37° C. The medium contained 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. One day before compound treatment, raw264.7 cells were seeded on a 6-well plate at 50,000 cells/well. The next day, the cells were pre-treated with compound CMDD-X, for 1 hour, by the concentrations, and then were treated with 100 ng/ml LPS for 24 hours, and the qRT-PCR was conducted. TNF-α and IL-6 are cytokines involved in the occurrence of atherosclerosis, and generated mainly in the macrophage to induce an inflammatory reaction and induce leukocytes, to thereby advance atherosclerosis. As the compound CMDD-X concentration increased, expressions of TNF-α and IL-6 genes significantly decreased (
The nitric oxide (NO) concentration in the medium of the raw264.7 macrophage treated in the experiment was measured. NO is generated in the macrophage or foam cell present in a lesion site where atherosclerosis is advanced. This causes oxidative stress in the blood to generate oxLDL, and is involved in the rupture of the atherosclerosis progression site. In addition, it may be seen from iNOS previously reported and the apoE-deficient mouse results that NO generated by iNOS functions as an important factor in the occurrence and progression of atherosclerosis (Detmers P A et. al., J. Immunol. 2000, 3430-3435). As a result of NO assay, as the compound CMDD-X concentration increased, expression of NO significantly decreased (
MCP-1, which is cytokine importantly involved in the atherosclerosis, is generated in the endotheliocyte to promote induction of monocytes into the inflammatory site and thus contributes to formation of atherosclerosis early lesions (Gerszten R E et al, NATURE, 1999, 718-723). HUVEC endotheliocytes were cultured in EGM-2 medium within a cell incubator containing 5% carbon dioxide at 37° C. One day before treatment with the compound CMDD-X, HUVEC endotheliocytes were seeded on a 6-well plate at 30,000 cells/well. The next day, the cells were pre-treated with compound CMDD-X, for 1 hour, by the concentrations, and then were treated with 10 ng/ml TNF-α for 6 hours, and the qRT-PCR was conducted. As a result of qRT-PCR, as the compound CMDD-X concentration increased, expression of MCP-1 significantly decreased (
VCAM-1, which is another important factor of atherosclerosis, mediates attachment of monocytes in the early first stage of atherosclerosis and is expressed at the edge of the lesion to be involved in the expansion of the lesion (Nakashima Y et al., Arterioscler Thromb Vasc Biol, 1998, 842-851). In order to verify efficacy of the compound CMDD-X on expression of the intercellular adhesion molecule gene (VCAM-1) in the HUVEC endotheliocytes and expression of protein, the samples were obtained by the same experiment and then qRT-PCR and Western experiments were conducted. As a result, expression of VCAM-1 was significantly decreased by the compound CMDD-X and expression of VCAM-1 protein was significantly decreased (
In the present example, in order to verify efficacy to prevent and treat Parkinson's Disease of the compound of Chemical Formula I according to the present invention in the Parkinson's Disease as one of neurodegenerative diseases, mice administered with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were used. The efficacy to prevent and treat the Parkinson's Disease was verified by administering the compound of Chemical Formula I (CMDD-X or Compound 7) to a Parkinson's Disease-induced disease model and then measuring exercise capacity of the mice. C57BL/6 mice (male, 12-week old) were used. The mice were adapted to a housing environment for 1 week while pellet type general diet was supplied, and then divided into 3 groups of 5 mice each to have similar body weights. Before MPTP administration, the compound CMDD-X was orally administered at a concentration of 5 mg/kg. After 3 hours, probenecid as MPTP efflux inhibitor was intraperitoneally administered to the mice at a concentration of 250 mg/kg, and, after 2 hours, 25 mg/kg of MPTP was intraperitoneally administered. Administration was conducted once per one day for a total of 5 days, and, after 3 days, exercise capacity of the mice was measured by using the Rotarod. The exercise capacity test was conducted by measuring the time while the mouse placed on the rod rotating at a predetermined speed was maintained on the rod without falling. The maximum 180 seconds were given, the speed was gradually increased, and the same break time was given. As a result of test, exercise capacity of the compound CMDD-X-administered group was recovered to a similar level to normal mice (
The compound of Chemical Formula I of the present invention (e.g., CMDD-X or Compound 7) may be orally or non-orally administered to mammals including human beings. The use of the compound is not limited to a particular formulation, and may be formulated into oral administration, drip liquid, tablet, trituration, liquid suppository, external application, patch, intravenous injection, powder, granule, sugar-coated tablet, capsule, pill, suspension, liquid, ampoule, injection, or the like, and may be applied to any other shape of medicine.
The tablet was prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 10 mg
Lactose 95 mg
hydroxypropylcellulose 3 mg
Calcium carboxymethylcellulose 8 mg
Magnesium stearate 0.5 mg
The powder was prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 15 mg
Lactose 20 mg
Starch 15 mg
Magnesium stearate: adequate amount
The hard capsule was prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 20 mg
Lactose 30 mg
Starch 25 mg
Talc 2 mg
Magnesium stearate adequate amount
Contents of the soft capsule were prepared by the conventional method while mixing the following components. The soft capsule was prepared by the conventional method using gelatin 132 mg, concentrated glycerin 50 mg, 70% D-sorbitol 6 mg, adequate amount of ethylvanillin as fragrance ingredient, and Carnauba Wax as a coating agent per one capsule.
Compound of Chemical Formula I (CMDD-X, Compound 7) 20 mg
Polyethyleneglycol 400 400 mg
Concentrated glycerin 50 mg
Purified water 35 mg
The suspension was prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 15 mg
Isomerized sugar 10 g
Sugar 25 mg
Calcium carboxymethylcellulose 50 mg
Lemony flavor adequate amount
Total preparation amount was adjusted to 100 ml by using purified water.
Each ampoule (2 ml) containing contents of the following components was prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 10 mg
Mannitol 180 mg
Sterile distilled water for injection 2974 mg
Disodium phosphate 26 mg
Ointments (ointment examples 1 to 3) were prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 0.5 wt %
White Petrolatum 70 wt %
Mineral oil 5 wt %
Polyoxylstearyl ether 5 wt %
Polyethylene glycol (ex.: PEG 400 or USP) 19.2 wt %
Butylhydroxyanisol 0.3 wt %
Compound of Chemical Formula I (CMDD-X, Compound 7) 0.5 wt %
White Petrolatum 70 wt %
Mineral oil 5 wt %
Polyoxylstearyl ether 5 wt %
Polyethylene glycol (ex.: PEG 400 or USP) 19.2 wt %
Butylhydroxyanisol 0.2 wt %
Paraben (ex.: Methylparaben or propylparaben) 0.1 wt %
Compound of Chemical Formula I (CMDD-X, Compound 7) 0.01 to 2 wt %
White Petrolatum 30 to 75 wt %
Mineral oil 2 to 10 wt %
Polyoxylstearyl ether 0.3 to 10 wt %
Polyethylene glycol (ex.: PEG 400 or USP) 2 to 45 wt %
Butylhydroxyanisol 0 wt % or 0.002 to 2.5 wt %
Paraben (ex.: methylparaben or propylparaben) 0 wt % or 0.01 to 1.5 wt %
Creams for external local use (cream examples 1 and 2) were prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 0.1 wt %
White Petrolatum 40 wt %
Mineral oil 11 wt %
Polyoxylstearyl ether 8.5 wt %
Propyleneglycol 18 wt %
Butylhydroxyanisol 0.02 wt %
Purified water adequate amount (22.38 wt %)
Compound of Chemical Formula I (CMDD-X, Compound 7) 0.01 to 3 wt %
White Petrolatum 30 to 75 wt %
Mineral oil 2 to 10 wt %
Polyoxylstearyl ether 0.3 to 10 wt %
Propyleneglycol 0.3 to 45 wt %
Butylhydroxyanisol 0.002 to 2.5 wt %
Paraben (ex.: Methylparaben or propylparaben) 0.01 to 3.5 wt %
Purified water appropriate amount (2 to 30 wt %)
The beverage was prepared to have a total volume of 100 mL by the conventional method while mixing an adequate amount of purified water with the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 2 mg
Citric acid 9 g
White sugar 9 g
Glucose 2.8 g
DL-malic acid 0.25 g
Purified water adequate amount
10-1. Preparation of Seasoning for Cooking
Seasoning for cooking for health promotion was prepared by using 0.001˜0.2 parts by weight of Compound of Chemical Formula I (CMDD-X, Compound 7).
10-2. Preparation of Flour Based Food
Food for health promotion was prepared by using the flour added with 0.001˜0.2 parts by weight of Compound of Chemical Formula I (CMDD-X, Compound 7) to make bread, cakes, cookies, crackers, and noodles.
10-3. Preparation of Dairy Products
Various dairy products such as butter, ice cream, milk powder, and baby food, were prepared by using milk added with 0.001˜0.2 parts by weight of Compound of Chemical Formula I (CMDD-X, Compound 7).
10-4. Preparation of Juice
Juice for health promotion was prepared by adding 0.001-0.2 parts by weight of Compound of Chemical Formula 1 (CMDD-X, Compound 7) to 1,000 m of juice of vegetable such as tomato or carrot or fruit such as apple, grape, orange, or pineapple.
1 kg of feedstuff for animal was prepared by the conventional method while mixing the following components.
Compound of Chemical Formula I (CMDD-X, Compound 7) 0.1 g
Casein 200 g
Corn starch 400 g
Dyestrose 130 g
Sugar 100 g
Cellulose 50 g
Soybean oil 70 g
Antioxidant (ex.: t-butylhydroquinone) 0.02 g
Inorganic substance (ex.: salt mix) 35 g
Vitamin (ex.: vitamin mix) 10 g
L-cystine 3 g
Choline bitartrate 2 g
The sesterterpene compound of Chemical Formula I according to the present invention has superior Nurr1 activation, and thus can control and maintain blood sugar, treat insulin-independent diabetes, and prevent the occurrence of diabetes complications. Further, the sesterterpene compound of Chemical Formula I according to the present invention can be used as an effective component of a composition for improving, treating, and preventing diabetes and diabetes complications (foot ulcer and renal failure) by improving insulin sensitivity through regulation of hormones associated with glucose metabolism and protecting pancreatic function to thereby control fasting blood sugar and prevent the occurrence of diabetes complications (foot ulcer and renal failure).
Further, the sesterterpene compound of Chemical Formula I according to the present invention can have a superior effect in inhibiting differentiation of adipocytes, and thus can be useful in improving, preventing, and treating obesity.
Further, the sesterterpene compound of Chemical Formula I according to the present invention may be used as an effective component of a composition for preventing, treating, and improving alcoholic, non-alcoholic, and viral fatty liver diseases by inhibiting the generation of fatty acids in the liver and promoting beta-oxidation by which the fatty acids are burn to release the heat to thereby significantly reduce fat accumulation in the liver.
Further, the sesterterpene compound of Chemical Formula I according to the present invention reduces low-density lipoprotein (LDL) cholesterol and inhibits expressions of inflammatory cytokines derived from macrophage and endotheliocyte, signaling proteins, and lipid biosynthesis enzymes, and thus can be used as an effective component of a composition for improving, treating, and preventing vascular diseases such as hyperlipidemia and atherosclerosis, alone or in a complex agent together with the existing LXR agonists that have been developed until the present time.
Further, the sesterterpene compound of Chemical Formula I according to the present invention that activates Nurr1 may be used alone as an effective component of a composition for improving, treating, and preventing brain disorders such as Parkinson's disease, schizophrenia, and manic-depression.
Further, the sesterterpene compound of Chemical Formula I according to the present invention may be used as an effective component of a composition for improving, treating, and preventing Alzheimer's disease in a complex agent together with the existing LXR agonists.
Number | Date | Country | Kind |
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10-2010-0087552 | Sep 2010 | KR | national |
10-2011-0020118 | Mar 2011 | KR | national |
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/KR2011/006638 | 9/7/2011 | WO | 00 | 5/24/2013 |