CN104130240A - Thioureathiophene-containing compound and application thereof - Google Patents
Thioureathiophene-containing compound and application thereof Download PDFInfo
- Publication number
- CN104130240A CN104130240A CN201410331189.2A CN201410331189A CN104130240A CN 104130240 A CN104130240 A CN 104130240A CN 201410331189 A CN201410331189 A CN 201410331189A CN 104130240 A CN104130240 A CN 104130240A
- Authority
- CN
- China
- Prior art keywords
- compound
- thioureido
- thiophenes
- medicine
- phenyl
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 0 CC(C)(CC1)CC(C)(C)Cc2c1c(C(O)=O)c(NC(NC(*)=O)=S)[s]2 Chemical compound CC(C)(CC1)CC(C)(C)Cc2c1c(C(O)=O)c(NC(NC(*)=O)=S)[s]2 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
- C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
- C07D333/52—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes
- C07D333/62—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to carbon atoms of the hetero ring
- C07D333/68—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
- C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
- C07D333/78—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems condensed with rings other than six-membered or with ring systems containing such rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a thioureathiophene-containing compound shown as a formula (I) and an application thereof as a sterol regulatory element binding protein small molecular conditioning agent. The compound has the capability of lowering the mitochondrial membrane potential, has good in-vitro and in-vivo lipid-decreasing effects, and can be taken as a lead compound for treating metabolic syndromes.
Description
Technical field
The invention belongs to medical technical field, relate to containing thioureido thiophenes and as the application of Sterol regulatory element binding protein (SREBP) small-molecule modulators.
Background technology
The prevalence of non-infectious chronics such as Jin Shinian China urban and rural residents hypertension, diabetes, obesity, hyperlipemia rise rapidly.End 2012, Chinese fat number has reached 1.2 hundred million left and right, accounts for 10% of total population.Obesity is that metabolism syndrome is the most also the most dangerous factor, and the weight index of normal population (BMI) should remain between 20-23.Studies show that, when people BMI surpasses 24, suffer from hypertensive danger and be the 3-4 of normal population doubly, the danger of suffering from diabetes be normal population 2-3 doubly; Over the danger of suffering from coronary heart disease at 29 o'clock, obviously increase; BMI breaks through at 30 o'clock, suffers from the risk of stating metabolism syndrome and will improve 10 times.
The core of metabolism syndrome is insulin resistant.It is the key character of metabolism syndrome that interior fat is piled up, and is also the major cause that causes insulin resistant.Think that at present visceral adipose is affected by genetic background, asian ancestry crowd has the advantages that fat is easily deposited in internal organ.At interior fat, pile up in individuality, the internal organs of first getting involved are livers.Too much the deposition of free fatty acids can cause fatty liver, and can cause that liver enzyme level raises, and the change of liver structure even occurs.And liver is one of vitals of lipogenesis, its building-up process is subject to the Regular Insulin of islet secretion and the feedback regulation of hyperglycemic-glycogenolytic factor.In liver, the dystopy of lipid deposition is the key link of metabolism syndrome morbidity, relevant to the generation of fatty liver, diabetes B and complication.In liver, Sterol regulatory element binding protein (Sterol Regulatory Element Binding Protein, SREBP) be the nuclear factor of a class key, can directly activate more than 30 genetic expression that participates in lipid acid, triglyceride level, cholesterol biosynthesis and picked-up, in lipid metabolism signal network, play important regulating and controlling effect.Studies show that in a large number, SREBP participates in the composition of insulin signaling pathway, simultaneously at the equal high expression level of hyperlipidaemia, insulin resistant, diabetes and Patients with Fatty Liver.The SREBP of take treats obesity as drug target becomes study hotspot day by day.
Chen Jie etc. are at rat nonalcoholic fatty liver (Nonalcoholic Fatty Liver Disease, NAFLD) in research, find, (the Liver X Receptor of liver X receptor after medicine rosiglitazone in treating, LXR), SREBP hypotype SREBP-1c expression level all obviously declines, and early intervention is better than intervening late period, and is dose-dependently.One is used for the treatment of the diabetes that cause due to insulin resistant N1,N1-Dimethylbiguanide medicine (Metformin), SREBP-1c level and lipid accumulation in ob/ob mouse can reduce liver (P.Pettinelli, et al.Bba-Mol Basis Dis, 2009,1792,1080-1086).In rats'liver primary cell, N1,N1-Dimethylbiguanide activates adenylic acid (AMP) activated protein kinase (AMP-activated Protein Kinase, AMPK), causes SREBP-1c and and the lipogenetic down regulation of gene expression in its downstream.The people such as Song Baoliang (J.J.Tang, et al.Cell Metab, 2011,13,44-56) by the luciferase reporter gene screening method based on SRE sequence, discovery natural product betulin is induced SREBP cracking activator (SREBP Cleavage-Activated Protein by specificity, SCAP) and insulin-induced gene (Insulin induced gene, Insig) interact, thereby SREBP/SCAP complex body is trapped in endoplasmic reticulum, and then reduces the generation of SREBP.Betulin can reduce the biosynthesizing of cholesterol and lipid acid in vitro; In vivo, betulin can improve food-induced obesity, reduces the content of tissue and lipids in serum, increases insulin sensitivity.In addition, betulin can significantly reduce atherosclerotic plaque area.As a SREBP micromolecular inhibitor, betulin can improve the whole disorders of lipid metabolism of obesity mice, has proved that suppressing SREBP regulates path may become the strategy of new treatment metabolic trouble.
Summary of the invention
One of object of the present invention is to propose a class containing thioureido thiophenes, and it has following structural formula (I):
In formula (I), R
1for alkyl, cycloalkyl, phenyl, aryl, thienyl or pyridyl; N=1 or 3.
Preferably, R
1for ethyl, amyl group, cyclohexyl, phenyl, methyl substituted phenyl, difluorophenyl, hydroxyl-substituted base, thienyl, pyridyl.
Formula of the present invention (I) is containing the preparation method of thioureido thiophenes, and reaction formula is as follows:
Wherein, R
1for alkyl, cycloalkyl, phenyl, aryl, thienyl or pyridyl; N=1 or 3.
The present invention also provides formula (I) containing the application in preparation treatment metabolic disease medicine as SREBP small-molecule modulators of thioureido thiophenes.Described metabolic disease comprises obesity, diabetes, fatty liver.The present invention also provides formula (I) containing the application in preparing the medicine that reduces medicine, the medicine of reduction body white lipid content, the medicine of regulation and control glucose metabolism genes of body weight gain or improve glucose-tolerant as SREBP small-molecule modulators of thioureido thiophenes.The present invention's application comprises that formula (I) is containing the application in reducing body weight gain as SREBP small-molecule modulators of thioureido thiophenes; The application of this compound in reducing body white lipid content; The application of this compound in the regulation and control of glucose metabolism genes; The application of this compound in the improvement of glucose-tolerant etc.Particularly, formula of the present invention (I) can reduce significantly body weight containing thioureido thiophenes and have a net increase of length, the obvious reduction of each main white adipose content and the improvement of glucose-tolerant in comitative aspect, can reduce fat with hypercholesterolemia and low-density lipoprotein/high-density lipoprotein (HDL) ratio, reduce and suffer from atherosclerotic risk, in addition obesity mice fatty liver is had some improvement, in liver, the content of triglyceride level and cholesterol obviously reduces, and liver heavily reduces.
The present invention also provides formula (I) containing thioureido thiophenes, to suppress the method for body weight gain, formula (I) reduces the method for body white lipid content containing thioureido thiophenes, formula (I) reduces the method for low-density lipoprotein/high-density lipoprotein (HDL) ratio containing thioureido thiophenes, and formula (I) is containing content, the heavy method of reduction liver of triglyceride level and cholesterol in thioureido thiophenes reduction liver.
In a specific embodiments, formula (I) can reduce the crucial synthetic enzyme of the interior SREBP-1c of liver and downstream synthetic fatty acid and triglyceride level containing thioureido thiophenes, as FAS, and ACS, ATP-CL transcribes.In a specific embodiments, formula (I) can make SREBP-2 transcriptional level lower containing thioureido thiophenes, and its target gene is as HMGCR, HMGCS, and the mRNA level of the synthetic cholesterol key enzymes such as SS is all lowered.The level of SREBP-1a is not subject to the impact of the compounds of this invention.
PPAR α is the main transcription factor of being responsible for regulation and control Fatty Acid Oxidation genes involved, and in a specific embodiments, after the compounds of this invention applies, this gene, in the level of liver organization, considerable change does not occur.
Longer chain fatty acid β-oxidation is the main path that animal obtains energy, long-chain acyl-CoA desaturase (LCAD) is the katalaze enzyme of longer chain fatty acid β-oxidation initial step, in a specific embodiments, apply after the compounds of this invention, but considerable change does not occur long-chain acyl-CoA desaturase, show that the compounds of this invention does not affect lipid oxidation process.
In a specific embodiments, glyconeogenesis key enzyme is as PEPCK, and G6Pase gene is significantly lowered after the compounds of this invention treatment.
PDC has limited pyruvic acid by complete oxidation or has been transformed into lipid acid after by PDK phosphorylation inactivation.Meanwhile, the activity that increases PDC by suppressing PDK is also the drug target for the treatment of diabetes, heart trouble and tumour, in a specific embodiments, applies the mRNA level that the compounds of this invention does not affect PDK4.
The present invention also provides formula (I) application in the regulation and control that reduce mitochondrial membrane potential in anoxic containing thioureido thiophenes.The present invention also provides formula (I) containing the impact of thioureido thiophenes on L6 inoblast mitochondrial membrane potential, L6 inoblast, add the compounds of this invention to cultivate, present the gentle ability that reduces mitochondrial membrane potential of the compounds of this invention, and showed certain concentration dependent.
Beneficial effect of the present invention also comprises, formula of the present invention (I) is novel compound containing thioureido thiophenes, there is different structure with prior art such as rosiglitazone, N1,N1-Dimethylbiguanide, betulin, rapamycin, trans-resveratrol etc., can be used as SREBP micromolecular inhibitor.The compounds of this invention in vitro and in body, all have good lipid-lowering effect, for example, obvious at DIO mouse lipid-lowering effect, can be used as the lead compound for the treatment of metabolic syndrome.
Accompanying drawing explanation
Fig. 1 represents that the compounds of this invention reduces the effect of L6 inoblast mitochondrial membrane potential.
Fig. 2 represent the compounds of this invention (compound 1, ZJ001) long term administration is on DIO Mouse Weight and the impact of ingesting; Wherein, A represents average food ration weekly; B represents the body weight change in administration 7 weeks; C represents the clean growth pattern of the body weight of administration after 7 weeks.
Fig. 3 represent the compounds of this invention (compound 1, ZJ001) long term administration is on the glycometabolic impact of DIO mouse; Wherein, A represents fasting plasma glucose.B represents the abdominal injection glucose-tolerant experiment of administration after 4 weeks.C represents area under glucose tolerance curve.
Fig. 4 represents the compounds of this invention (compound 1, the ZJ001) regulation and control to glycolipid metabolism gene after long-term treatment; Wherein, A-C represents fat synthesis related gene; D represents lipid oxidation genes involved; E represents carbohydrate metabolism genes involved.
Embodiment
In conjunction with following specific embodiments and the drawings, the present invention is described in further detail, and protection content of the present invention is not limited to following examples.Do not deviating under the spirit and scope of inventive concept, variation and advantage that those skilled in the art can expect are all included in the present invention, and take appending claims as protection domain.Implement process of the present invention, condition, reagent, experimental technique etc., except the content of mentioning specially below, be universal knowledege and the common practise of this area, the present invention is not particularly limited content.Each embodiment raw material used is commercially available analytical pure chemical.
Embodiment 1 the compounds of this invention 1 (ZJ001, R
1=Ph, n=1) Preparation and characterization
Compound a 1 (1.30g, 8mmol) and b1 (2.02g, 8mmol) are dissolved in 20ml acetonitrile, heating reflux reaction 5h, and cooling rear suction filtration, washing with alcohol twice for filter cake, obtains yellow powder shape compound c1.
Compound c1 (0.42g, 1mmol) is placed in 5ml CF
3in COOH, stirring at room reaction 1h, removes excessive CF
3cOOH, then adds 10ml isopropyl ether, and after fully stirring, suction filtration, dry, obtains yellow solid compound 1 (ZJ001).Yield 62%,
1hNMR (500MHz, DMSO-d
6): δ 14.79 (s, 1H), 13.22 (s, 1H), 11.70 (s, 1H), 7.96 (d, J=7.5Hz, 2H), 7.67-7.64 (m, 1H), 7.55-7.52 (m, 2H), 2.76 (s, 2H), 2.64 (s, 2H), 1.73 (s, 4H);
13cNMR (125MHz, DMSO-d
6): δ 174.25,166.75,165.62,145.49,133.04,132.16,131.70,128.73 (2C), 128.43 (2C), 127.44,116.79,25.82,23.71,22.51,22.35; HRMS (ESI): calcd for C
17h
16n
2o
3s
2na:383.0500[M+Na]
+, found:383.0543[M+Na]
+; M.p.227 ℃.
Embodiment 2 the compounds of this invention 2 (R
1=o-MeC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with o-methyl-phenyl-.Make compound 2: yield 89%,
1h NMR (400MHz, DMSO-d
6) δ 14.71 (s, 1H), 11.82 (s, 1H), 7.51 (d, J=7.6Hz, 1H), 7.44 (t, J=7.5Hz, 1H), 7.34-7.26 (m, 2H), 2.77 (s, 2H), 2.65 (s, 2H), 2.40 (s, 3H), 1.75-1.73 (m, 4H).
13c NMR (101MHz, DMSO-d
6) δ 174.09,168.78,165.69,145.37,136.01,133.93,131.56,130.87,130.55,128.14,127.35,125.48,116.60,25.73,23.61,22.43,22.27,19.40; HRMS (ESI) calcd for C
18h
18n
2o
3s
2na:397.0651[M+Na]
+, found:397.0667[M+Na]
+; M.p.236-237 ℃.
Embodiment 3 the compounds of this invention 3 (R
1=m-MeC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with an aminomethyl phenyl.Make compound 3: yield 86%,
1h NMR (400MHz, DMSO-d
6) δ 14.76 (s, 1H), 11.62 (s, 1H), 7.81 (s, 1H), 7.75 (d, J=7.6Hz, 1H), 7.48-7.40 (m, 2H), 2.77 (s, 2H), 2.64 (s, 2H), 2.39 (s, 3H), 1.74-1.74 (m, 4H).
13c NMR (101MHz, DMSO-d
6) δ 174.20,166.74,165.52,145.46,137.75,133.61,131.96,131.63,129.11,128.31,127.35,125.84,116.71,25.75,23.62,22.43,22.26,20.77; HRMS (ESI) calcd for C
18h
18n
2o
3s
2na:397.0651[M+Na]
+, found:397.0658[M+Na]
+; M.p.233-234 ℃.
Embodiment 4 the compounds of this invention 4 (R
1=p-MeC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with p-methylphenyl.Make compound 4: yield 87%;
1hNMR (500MHz, DMSO-d
6); δ 14.78 (s, 1H), 13.23 (s, 1H), 11.62 (s, 1H), 7.89 (d, J=8.0Hz, 2H), 7.34 (d, J=8.0Hz, 2H), 2.77 (s, 2H), 2.65 (s, 3H), 2.40 (s, 3H), 1.56-1.53 (m, 4H);
13cNMR (125MHz, DMSO-d
6); δ 174.07,170.47, and 165.22,145.12,131.49,127.26,116.57,25.64,23.56,23.48,22.39,22.21; HRMS (ESI) calcd for C
18h
18n
2o
3s
2na:397.0651[M+Na]
+, found:397.0667[M+Na]
+; M.p.229-231 ℃.
Embodiment 5 the compounds of this invention 5 (R
1=o-FC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with adjacent fluorophenyl.Make compound 5: yield 84%;
1h NMR (400MHz, DMSO-d
6) δ 14.64 (s, 1H), 13.26 (s, 1H), 11.86 (s, 1H), 7.71-7.64 (m, 2H), 7.35 (m, 2H), 2.77 (s, 2H), 2.65 (s, 2H), 1.74 (s, 4H).
13c NMR (101MHz, DMSO-d
6) δ 173.51,165.58,163.79,145.22,134.07,133.98,131.70,130.40,127.54,124.53,116.79,116.24,116.02,25.74,23.63,22.42,22.25; HRMS (ESI) calcd for C
17h
15n
2o
3s
2naF:401.0406[M+Na]
+, found:401.0418[M+Na]
+m.p.240-241 ℃.
Embodiment 6 the compounds of this invention 6 (R
1=m-FC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with a fluorophenyl.Make compound 6: yield 83%,
1h NMR (400MHz, DMSO-d
6) δ 14.73 (s, 1H), 11.80 (s, 1H), 7.81-7.78 (m, 2H), 7.62-7.56 (m, 1H), 7.51 (td, J=9.3,2.08Hz, 1H), 2.77 (s, 2H), 2.65 (s, 2H), 1.75-1.73 (m, 4H).
13c NMR (101MHz, DMSO-d
6) δ 173.98,165.62 (2C), 145.25,134.41,134.34,131.65,130.56,130.49,127.45,116.74,115.68,115.45,25.74,23.61,22.42,22.26; HRMS (ESI) calcd for C
17h
15n
2o
3s
2naF:401.0406[M+Na]
+, found:401.0424[M+Na]
+; M.p.233-234 ℃.
Embodiment 7 the compounds of this invention 7 (R
1=p-FC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with fluorophenyl.Make compound 7: yield 85%,
1hNMR (500MHz, DMSO-d
6) δ 14.74 (s, 1H), 13.25 (br, 1H), 11.74 (s, 1H), 8.04 (t, J=6.25Hz, 2H), 7.34 (t, J=8.5Hz, 2H), 2.75 (s, 2H), 2.62 (s, 2H), 1.71 (s, 4H);
13cNMR (125MHz, DMSO-d
6) δ 174.22,165.69,165.61,163.91,145.34,131.78,131.70,131.67,128.62,127.43,116.76,115.52,115.34,25.80,23.67,22.48,22.32; HRMS (ESI) calcd for C
17h
15n
2o
3s
2naF:401.0406[M+Na]
+, found:401.0398[M+Na]
+; M.p.221-222 ℃.
Embodiment 8 the compounds of this invention 8 (R
1=o-OHC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with o-hydroxy-phenyl.Make compound 8: yield 33%,
1h NMR (400MHz, DMSO-d
6) δ 14.77 (s, 1H), 11.78 (s, 1H), (7.97 d, J=7.4Hz, 1H), (7.53 t, J=7.1Hz, 1H), (7.04 t, J=7.6Hz, 2H), (2.77 s, 2H), 2.65 (s, 2H), 1.74 (s, 4H) .HRMS (ESI) calcd for C
17h
16n
2o
4s
2na:399.0449[M+Na]
+, found:399.0443[M+Na]
+m.p.232-233 ℃.
Embodiment 9 the compounds of this invention 9 (R
1=m-OHC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with a hydroxy phenyl.Make compound 9: yield 24%,
1h NMR (500MHz, DMSO-d
6) δ 13.25 (br, 1H), 12.33 (s, 1H), 10.02 (s, 1H), 7.41 (t, J=7.9Hz, 1H), 7.31 (s, 1H), 7.05 (d, J=7.5Hz, 1H), 2.75 (s, 2H), 2.64 (s, 2H), 1.74 (s, 4H).
13c NMR (101MHz, DMSO-d
6) δ 174.15,166.63,165.49,157.26,145.28,133.41,131.65,129.50,127.33,120.03,119.18,116.74,115.24,25.74,23.64,22.45,22.29.HRMS (ESI) calcd for C
17h
16n
2o
4s
2na:399.0449[M+Na]
+, found:399.0446[M+Na]
+; M.p.228-229 ℃.
Embodiment 10 the compounds of this invention 10 (R
1=p-OHC
6h
4, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with p-hydroxybenzene.Make compound 10: yield 21%,
1h NMR (400MHz, DMSO-d
6) δ 14.78 (s, 1H), 13.18 (br, 1H), 11.40 (s, 1H), 10.41 (s, 1H), 7.89 (d, J=8.7Hz, 2H), 6.86 (d, J=8.8Hz, 2H), 2.76 (s, 2H), 2.64 (s, 2H), 1.74-1.73 (m, 4H).
13c NMR (101MHz, DMSO-d
6) δ 174.54,166.02,165.43,162.09,145.35,131.59,131.17 (2C), 127.28,122.07,116.63,115.11 (2C), 25.74,23.62,22.44,22.29; HRMS (ESI) calcd for C
17h
16n
2o
4s
2na:399.0449[M+Na]
+, found:399.0451[M+Na]
+m.p.251-252 ℃.
Embodiment 11 the compounds of this invention 11 (R
1=2-thienyl, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with 2-thienyl.Make compound 11: yield 40%,
1h NMR (500MHz, DMSO-d
6) δ 14.68 (s, 1H), 13.21 (s, 1H), 11.65 (s, 1H), 8.35 (d, J=2.7Hz, 1H), 8.03 (d, J=4.5Hz, 1H), 7.25 (t, J=4.0Hz, 1H), 2.76 (s, 2H), 2.64 (s, 2H), 1.73 (s, 4H);
13cNMR (125MHz, DMSO-d
6) δ 173.72,165.52,160.49,145.83,136.76,135.15,132.63,131.72,128.73,127.33,116.86,25.81,23.72,22.53,22.36; HRMS (ESI) calcd for C
15h
14n
2o
3s
3na:389.0064[M+Na]
+, found:389.0015[M+Na]
+; M.p.210-211 ℃.
Embodiment 12 the compounds of this invention 12 (R
1=4-pyridyl, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with 4-pyridyl.Make compound 12: yield 90%;
1h NMR (500MHz, DMSO-d
6) δ 13.50 (br, 1H), 12.43 (s, 1H) 8.86 (dd, J=1.3,4.6Hz, 2H), 7.79 (dd, J=1.4,4.6Hz, 2H), 2.75 (s, 2H), 2.65 (s, 2H), 1.75-1.73 (m, 4H);
13c NMR (101MHz, DMSO-d
6) δ 167.44,160.97,150.81 (2C), 145.54,139.26,131.38,126.77,120.73 (2C), 113.39,25.79,23.76,22.48,22.17; HRMS (ESI) calcd for C
16h
15n
3o
3s
2na:384.0453[M+Na]
+, found:384.0447[M+Na]
+; M.p.233 ℃.
Embodiment 13 the compounds of this invention 13 (R
1=3-pyridyl, Preparation and characterization n=1)
With reference to method shown in embodiment 1, wherein a1 " phenyl " replaces with 3-pyridyl.Make compound 13: yield 84%,
1h NMR (500MHz, DMSO-d
6) δ 12.36 (s, 1H), 9.05 (d, J=1.3,1H), 8.82 (d, J=4.05Hz, 1H), 8.26 (d, J=7.8Hz, 1H) 7.66 (dd, J=4.95,7.8Hz, 1H), 2.71 (s, 2H), 2.61 (s, 2H), 1.71 (s, 4H); HRMS (ESI) calcd for C
16h
15n
3o
3s
2na:384.0453[M+Na]
+, found:384.0442[M+Na]
+; M.p.242 ℃.
Embodiment 14 the compounds of this invention 14 (R
1=Ph, n=3) Preparation and characterization
With reference to the method shown in embodiment 1, wherein b1 " cyclohexyl " replaces with ring octyl group.Make compound 14: yield 86%,
1hNMR (500MHz, DMSO-d
6): δ 14.72 (s, 1H), 13.65 (s, 1H), 11.60 (s, 1H), 7.97 (d, J=7.5Hz, 2H), 7.67-7.64 (m, 1H), 7.55-7.52 (m, 2H), 2.93 (t, J=6.0Hz, 2H), 2.75 (t, J=6.0Hz, 2H), 1.61 (s, 4H), 1.44 (s, 2H), 1.24 (s, 2H);
13cNMR (125MHz, DMSO-d
6): δ 181.36,176.29,175.11,154.15,143.76,142.54,141.57,140.11,138.20 (2C), 137.91 (2C), 126.71,41.63,39.22,35.49,35.40,34.69,34.05; HRMS (ESI): calcd for C
19h
20n
2o
3s
2na:411.0813[M+Na]
+, found:411.0796[M+Na]
+; M.p.211 ℃.
Embodiment 15 the compounds of this invention 15 (R
1=ethyl, Preparation and characterization n=1)
With reference to method shown in embodiment 1, wherein a1 " phenyl " replaces with ethyl.Make compound 15: yield 76%,
1h NMR (400MHz, DMSO-d
6) δ 14.52 (s, 1H), 13.02 (br, 1H), 11.49 (s, 1H), 2.74 (s, 2H), 2.62 (s, 2H), 2.45 (q, J=6.8Hz, 2H), 1.72 (s, 4H), 1.05 (t, J=6.6Hz, 3H).
13c NMR (101MHz, DMSO-d
6) δ 174.16,174.10,165.26,145.17,131.54,127.32,116.57,28.93,25.70,23.59,22.43,22.27,8.53; HRMS (ESI) calcd for C
13h
16n
2o
3s
2na:335.0495[M+Na]
+, found:335.0511[M+Na]
+; M.p.230 ℃.
Embodiment 16 the compounds of this invention 16 (R
1=n-pentyl, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with n-pentyl.Make compound 16: yield 74%,
1h NMR (400MHz, DMSO-d
6) δ 14.52 (s, 1H), 13.12 (s, 1H), 11.48 (s, 1H), 2.74 (s, 2H), 2.62 (s, 2H), 2.43 (t, J=7.1Hz, 2H), 1.72 (s, 4H), 1.55 (s, 2H), 1.27 (s, 6H), 0.86 (s, 3H).
13c NMR (101MHz, DMSO-d
6) δ 174.14,173.4,165.29,145.17,131.54,127.34,116.59,35.54,30.90,28.10,25.70,24.19,23.60,22.43,22.27,21.89,13.88; HRMS (ESI) calcd for C
16h
22n
2o
3s
2na:377.0964[M+Na]
+, found:377.0966[M+Na]
+; M.p.237 ℃.
Embodiment 17 the compounds of this invention 17 (R
1=cyclohexyl, Preparation and characterization n=1)
With reference to the method shown in embodiment 1, wherein a1 " phenyl " replaces with cyclohexyl.Make compound 17:
1h NMR (400MHz, DMSO-d
6) δ 14.53 (s, 1H), 12.97 (br, 1H), 11.48 (s, 1H), 2.72 (s, 2H), 2.61 (s, 2H), 1.78-1.64 (m, 9H), 1.43-1.06 (m, 6H).
13c NMR (101MHz, DMSO) δ 176.21,174.38,165.26,145.13,131.51,127.30,116.55,43.65,28.55 (2C), 25.69,25.16,24.91 (2C), 23.57,22.40,22.24.HRMS (ESI) calcd for C
17h
22n
2o
38
2na:389.0964[M+Na]
+, found:389.0998[M+Na]
+; M.p.238 ℃.
The impact of embodiment 18 the compounds of this invention on L6 inoblast mitochondrial membrane potential
Cell cultures
L6 inoblast is containing 10% foetal calf serum, and 100kU/L penicillin, in the DMEM in high glucose substratum of 100mg/L Streptomycin sulphate, is placed in CO
2incubator (37 ℃, 5%CO
2, 95% air) cultivate, to density 60% left and right, go down to posterity.With 6 * 10
4/ mL concentration accesses (first day) in corresponding Tissue Culture Plate or Tissue Culture Dish, cultivates 24h to 80% adherent.Cell breaks up in the DMEM in high glucose substratum that contains 2% foetal calf serum, penicillin 100kU/L, Streptomycin sulphate 100mg/L, and every 48h changes liquid once.More than 70% be divided into muscle cell to the 7th day inoblast.
Mitochondrial membrane potential detects
1) L6 inoblast 6000/well (HG-DMEM, 10% foetal calf serum), is inoculated in blackboard clear bottom 96 orifice plates,
When density reaches 70%, change liquid differentiation (HG-DMEM, 2% foetal calf serum), break up after five days and can be used for testing;
2) at 100 μ L, containing in the HG-DMEM of 2% foetal calf serum, according to 100x, add corresponding compound (being respectively compound 1～compound 17 that previous embodiment 1～17 makes), discard the old substratum in 96 orifice plates, add the above-mentioned medium treatment certain hour (according to requirement of experiment) containing compound;
3) after finishing, compound treatment adds HG-DMEM (containing 2% foetal calf serum) the 100 μ L containing JC-1.96 orifice plate lucifuges are placed in 37 ℃ of incubator reaction required times;
4) nutrient solution in 96 orifice plates is all dried, on thieving paper, place a moment, then use Krebs-Ringer phosphate HEPES (KRPH) solution to wash three times, each 200 μ L.Last drying after solution, is inverted in thieving paper by 96 orifice plates and pats several times, adds 100 μ L KRPH solution to be detected;
5) detecting instrument Flexstation II384, red fluorescence ex/em530nm/580nm, green fluorescence ex/em485nm/530nm, reads plate mode well scan, reads altogether 9 regions.
6) data processing, derives respectively the mean value of red fluorescence and green fluorescence, deducts corresponding blank value, and the ratio of last red fluorescence and green fluorescence can represent the state of mitochondrial membrane potential.
7) positive control CCCP, working concentration is 10 μ M, be 30 minutes action time.
The compounds of this invention the results are shown in Figure 1 to the impact of L6 inoblast mitochondrial membrane potential, as can be seen from the results, the compounds of this invention 1,2,3,5,6,8,11,16,17 has all shown the ability of gentle reduction mitochondrial membrane potential, and presents certain concentration dependent.
Chronic pharmacodynamic evaluation in embodiment 19 representation compound compound 1 (ZJ001) bodies
Male wild-type C57BL/6J mouse is purchased from the Si Laike laboratory animal of Shanghai, and An Ping center, institute of materia medica, Bing You Shanghai is responsible for daily SPF level and is raised, and raising temperature is 22 ± 2 ℃, and humidity is 35 ± 5%, circulation in 12 hours round the clock.With every cage 4-5 mouse, raise, drink water at random and search for food, under starvation conditions, remove food but keep drinking-water.Wild-type male C 57 BL/6 J mouse started to give high lipid food from the 6th week, gave continuously 8 weeks, and the normal feed of simultaneously establishing coupling in age in week, gender matched gives group.Each treated animal twice of gastric infusion every day of dosage (ZJ001,15mg/kg) according to the rules, monitors food and body weight every day subsequently; Survey weekly fasting plasma glucose and random blood sugar; Within the 4th week, measure glucose tolerance test; Within the 5th week, afterbody is got blood, centrifuging and taking supernatant, and-80 degree are preserved, for the detection of blood plasma index; Within the 7th week, de-neck is put to death, and has got respectively liver, kidney, stomach fat, subcutaneous lipids, perirenal fat, leg muscle and abdominal muscles, and has weighed, and after liquid nitrogen flash freezer, is stored in-80 ℃ of cryogenic refrigerators and stores stand-by.The relevant detailed rules and regulations of Shanghai medicine institute experimentation on animals working specification are all followed in all mouse experiment operations.
The mensuration of plasma metabolite
Plasma high density lipoprotein level, low-density lipoprotein, triglyceride level and cholesterol are by each kit measurement of Zhangjiang biotechnology company of Shanghai Fudan University, blood plasma free fatty acid is by NEFA kit measurement (Wako Chemicals, Japan), plasma insulin is by ELISA kit measurement (Linco Research, St Louis, MO), blood sugar is measured (Abbott Laboratories) by Free-Style blood glucose meter.
The mensuration of abdominal muscles and liver tg and cholesterol
The mensuration of abdominal muscles and liver tg adopts the Folch method of standard, first take abdominal muscles and organize 40-50mg or liver organization 30-40mg, at chloroform: in (volume ratio is 2:1) after homogenate, spend the night with gyroscope suspendible under room temperature in methanol solution.Add concuss after the sodium-chlor of 2ml0.6%, then at 2000rpm centrifugal 10 minutes.The organic phase of taking out lower floor is placed in clean Glass tubing, is placed in stink cupboard, 45 ℃ of heat dryings.After dry, use 1-2ml anhydrous alcohol solution, then measure by triglyceride level detection kit and Cholesterol Kit.
Glucose-tolerant experiment
DIO mouse processes after 4 weeks to medicine (ZJ001,15mg/kg), first hungry 6 hours, every mouse with 2g/kg abdominal injection glucose after, at 0,15,30,60,90,120 minute, measure tail vein sugar value, and area AUC under calculated curve.
Real-time fluorescence quantitative PCR
1) preparation of sample: take the liver organization of 50mg left and right, add after 1ml TRIzol, grind with electric homogenizer.
2) the total mRNA of extracting and cDNA reversion: with extracting and the reversion of cell sample.
3) quantitative PCR: use the SYBR Green PCR test kit of Takara company, take β-actin as internal reference, carry out the relative of gene
Quantitatively.By Δ Δ CT method, carry out data analysis.
Fig. 2 represents that the compounds of this invention 1 (ZJ001) long term administration is on DIO Mouse Weight and the impact of ingesting.Wherein, Fig. 2 A represents average food ration weekly.Fig. 2 B represents the body weight change in administration 7 weeks.Fig. 2 C represents the clean growth pattern of the body weight of administration after 7 weeks.DIO mouse twice gavage every day gives Vehicle (0.5% methocel solution) and compound 1 (ZJ001) (15mg/kg), normal diet group gives Vehicle and treats 7 weeks as system negative control, monitors body weight and food consumption quantity every day.Result is all treated to mean ± SE (n=6-8).* P < 0.05, * * P < 0.01 (than HF-Veh).
As can be seen from Figure 2, the C57BL/6J mouse in 6 week age presents obesity, insulin resistant symptom after high lipid food is fed 8 weeks.With 15mg/kg/day oral dose administration every day after seven weeks, compound 1 is not in the situation that affecting DIO mouse and ingesting, as shown in Figure 2 A, body weight increases and slows down compared with control group (HF-Veh group), as shown in Figure 2 B, the length that has a net increase of of body weight has obvious reduction (49%) compared with control group, as shown in Figure 2 C.
After compound 1 treatment, respectively organize organ index and the plasma parameters of mouse, as shown in table 1.Consistent with body weight change, administration group represents epididymis white adipose, kidney week white adipose and represent that the weight of the inguinal region white adipose tissue of subcutaneous lipids also declines to a great extent by (table 1), especially perirenal fat and the subcutaneous lipids of internal organ white adipose.Wherein perirenal fat weight ratio has declined 23%, and subcutaneous lipids weight ratio has declined 42%.Fatty tissue is as the main place of depot fat, and the chronic rear body fat of compound 1 that gives obviously reduces.
High fat diet can improve suffers from atherosclerotic risk.One is using the ratio of cholesterol in blood plasma and low-density lipoprotein/high-density lipoprotein (HDL) as two atherosis indexs of prediction chronic arterial, and the level of high fat diet group mouse is the twice above (table 1) of normal diet group.In compound 1 administration group, these two indexs have reduced respectively 16% and 24%.Indication compound 1 can reduce obesity mice and suffer from atherosclerotic risk.Compound 1 can significantly reduce the accumulation of triglyceride level and cholesterol in the liver that high fat diet causes, and liver heavily has remarkable reduction, shows that compound 1 can improve fatty liver (table 1).
Fig. 3 represents the compounds of this invention, and (compound 1, ZJ001) long term administration is on the glycometabolic impact of DIO mouse.Wherein, A. fasting plasma glucose.B. the abdominal injection glucose-tolerant experiment of administration after 4 weeks.C. area under glucose tolerance curve.Result is all treated to mean ± SE. (n=6-8).* P < 0.05, * * P < 0.01, * * * p < 0.001 (with respect to HF-Veh).
In administration, after 4 weeks, compound 1 is on not impact of the hungry blood sugar level of DIO mouse, as shown in Figure 3A; Inject 2g/kg glucose simultaneously and evaluate the impact of ZJ001 on sugar tolerance, result shows that compound 1 can significantly reduce the blood sugar of 15min and 60min, area under glucose tolerance curve is added up, compared with control group, reduced by 13%, show that compound 1 has obviously improved glucose-tolerant ability, as shown in Fig. 3 B and 3C.
Fig. 4 represents the compounds of this invention (compound 1, the ZJ001) regulation and control to glycolipid metabolism gene after long-term treatment.Wherein, A-C. fat synthesis related gene.D. lipid oxidation genes involved.E. carbohydrate metabolism genes involved.Result is all treated to mean ± SE. (n=6-8).*P＜0.05，**P＜0.01，***p＜0.001。
Compound 1 can reduce the crucial synthetic enzyme of the interior SREBP-1c of liver and downstream synthetic fatty acid and triglyceride level, as FAS, and ACS, ATP-CL transcribes.Equally, SREBP-2 transcriptional level is also lowered, and its target gene is as HMGCR, HMGCS, and the mRNA level of the synthetic cholesterol key enzymes such as SS is all lowered.The level of SREBP-1a is not subject to the impact of ZJ001, as shown in Fig. 4 A-C.PPAR α is the main transcription factor of being responsible for regulation and control Fatty Acid Oxidation genes involved, and after compound 1 treatment, this gene, in the level of liver organization, considerable change does not occur.Longer chain fatty acid β-oxidation is the main path that animal obtains energy, long-chain acyl-CoA desaturase (LCAD) is the katalaze enzyme of longer chain fatty acid β-oxidation initial step, but considerable change does not occur yet for it, prompting compound 1 does not affect lipid oxidation process, as shown in Figure 4 D.Glyconeogenesis key enzyme is as PEPCK, and G6Pase gene is significantly lowered after compound 1 treatment.PDC has limited pyruvic acid by complete oxidation or has been transformed into lipid acid after by PDK phosphorylation inactivation.Meanwhile, the activity that increases PDC by suppressing PDK is also the drug target for the treatment of diabetes, heart trouble and tumour, but compound 1 does not affect the mRNA level of PDK4, as shown in Figure 4 E.
Above result shows, after the obesity mice of compound 1 (representation compound of the compounds of this invention) long-term treatment diet induced, its body weight gain is slack-off, has a net increase of longly significantly to reduce; White adipose tissue weight significantly reduces.In blood plasma, the content of cholesterol and the ratio of LDL/HDL significantly decline, and have reduced obesity mice and have suffered from atherosclerotic risk; After compound 1 long term administration, the liver weight of obesity mice reduces, and cholesterol and content of triglyceride reduce, and indication compound 1 can improve fatty liver to a certain extent.Compound 1 has also strengthened the glucose-tolerant ability of obesity mice, improves carbohydrate metabolism disturbance.
To sum up, the present invention series small molecules in vitro and in body, all obtain good lipid-lowering effect, can be used as the lead compound for the treatment of metabolic syndrome.
Table 1 is respectively organized organ index and the plasma parameters of mouse after ZJ001 chronic treatment
Every group of data that comprise 6-8 mouse, every group of data present with mean value ± standard error.Mouse forms obese model after high fat is raised 8 weeks, then uses ZJ001 oral administration 7 weeks, and dosage is 15mg/kg/day.* p < 0.05, * * p < 0.01, and * * * p < 0.001 is the blank administration group with respect to high fat.ND, represents not detect result.
Claims (6)
1. containing a thioureido thiophenes, it is characterized in that, its structure is suc as formula shown in (I):
In formula (I), R
1for alkyl, cycloalkyl, phenyl, aryl, thienyl or pyridyl; N=1 or 3.
2. as claimed in claim 1 containing thioureido thiophenes, it is characterized in that R
1for ethyl, amyl group, cyclohexyl, phenyl, methyl substituted phenyl, difluorophenyl, hydroxyl-substituted base, thienyl, pyridyl.
3. as claimed in claim 1 containing the application in preparation treatment metabolic disease medicine as SREBP small-molecule modulators of thioureido thiophenes.
4. application as claimed in claim 3, is characterized in that, described metabolic disease comprises obesity, diabetes, fatty liver.
As claimed in claim 1 containing thioureido thiophenes as SREBP small-molecule modulators preparation reduce body weight gain medicine, preparation reduce medicine, the medicine of preparation regulation and control glucose metabolism genes of body white lipid content or improve the application in the medicine of glucose-tolerant.
6. the application in the regulation and control that reduce mitochondrial membrane potential in anoxic containing thioureido thiophenes as claimed in claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410331189.2A CN104130240A (en) | 2014-07-11 | 2014-07-11 | Thioureathiophene-containing compound and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410331189.2A CN104130240A (en) | 2014-07-11 | 2014-07-11 | Thioureathiophene-containing compound and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104130240A true CN104130240A (en) | 2014-11-05 |
Family
ID=51803134
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410331189.2A Pending CN104130240A (en) | 2014-07-11 | 2014-07-11 | Thioureathiophene-containing compound and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104130240A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110437140A (en) * | 2019-07-16 | 2019-11-12 | 中国人民解放军总医院 | It is a kind of inhibit SREBP-1 target spot compound and its application |
-
2014
- 2014-07-11 CN CN201410331189.2A patent/CN104130240A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 杨玲玲: "线粒体调节剂C2的发现及作用机制研究", 《中国优秀硕士学位论文全文数据库（医药卫生科技辑）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110437140A (en) * | 2019-07-16 | 2019-11-12 | 中国人民解放军总医院 | It is a kind of inhibit SREBP-1 target spot compound and its application |
| CN110437140B (en) * | 2019-07-16 | 2021-08-03 | 中国人民解放军总医院 | Compound for inhibiting SREBP-1 target and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhao et al. | Structure-specific effects of short-chain fatty acids on plasma cholesterol concentration in male Syrian hamsters | |
| Feng et al. | ATP-citrate lyase (ACLY) in lipid metabolism and atherosclerosis: an updated review | |
| Kamanna et al. | Mechanism of action of niacin | |
| Yang et al. | Role of bone morphogenetic protein‐9 in the regulation of glucose and lipid metabolism | |
| Hasheminezhad et al. | A mechanistic insight into the biological activities of urolithins as gut microbial metabolites of ellagitannins | |
| Qi et al. | Effects of Morus root bark extract and active constituents on blood lipids in hyperlipidemia rats | |
| Xiong et al. | Antidiabetic activities of entagenic acid in type 2 diabetic db/db mice and L6 myotubes via AMPK/GLUT4 pathway | |
| US10370400B2 (en) | 4-methylumbelliferone derivatives for treatment for immune modulation | |
| Huang et al. | High-fructose and high-fat feeding correspondingly lead to the development of lysoPC-associated apoptotic cardiomyopathy and adrenergic signaling-related cardiac hypertrophy | |
| Lv et al. | AMP-kinase pathway is involved in tumor necrosis factor alpha-induced lipid accumulation in human hepatoma cells | |
| Weng et al. | Caffeic acid phenylethyl amide protects against the metabolic consequences in diabetes mellitus induced by diet and streptozocin | |
| KR20140125117A (en) | Composition for Preventing or Treating Hyperproliferative Vascular Disorders Comprising Carbanilide Derivatives as Active Ingredient | |
| CN101209257B (en) | Medical use of baicalin and scutellarein derivatives | |
| US11278518B2 (en) | Methods of treatment using 4-methylumbelliferone and derivatives thereof | |
| Li et al. | Liver cyclophilin D deficiency inhibits the progression of early NASH by ameliorating steatosis and inflammation | |
| Xu et al. | Berberine inhibits gluconeogenesis in skeletal muscles and adipose tissues in streptozotocin-induced diabetic rats via LKB1-AMPK-TORC2 signaling pathway | |
| CN102451178A (en) | Application of dihydrofuran-2-ketone compounds in preparing medicament for resisting diabetes mellitus and glucose and lipid metabolism | |
| CN110960535A (en) | Application of 18 β -glycyrrhetinic acid in preparation of medicines for treating diseases related to lipid metabolism disorder | |
| CN104130240A (en) | Thioureathiophene-containing compound and application thereof | |
| CN106456594A (en) | Ppar gamma activating agent | |
| An et al. | Physiological characterization of a novel PPAR pan agonist, 2-(4-(5, 6-methylenedioxybenzo [d] thiazol-2-yl)-2-methylphenoxy)-2-methylpropanoic acid (MHY2013) | |
| TW201731557A (en) | Beneficial effect of Bidens pilosa on fat decrease and muscle increase | |
| CN106397207B (en) | Tree ketonic acid A structural analogue, composition thereof and application thereof in medicines | |
| CN101006995A (en) | Application of isosteviol in pharmacy | |
| WO2014029360A1 (en) | Amino-substituted rutaecarpin analog and synthesis method and use thereof in preparation of anti-obesity drug |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20141105 |