WO2016138132A1 - Inhibiteurs de la dipeptidyl-peptidase iv (dpp4), procédés et compositions pour supprimer une inflammation des tissus adipeux - Google Patents

Inhibiteurs de la dipeptidyl-peptidase iv (dpp4), procédés et compositions pour supprimer une inflammation des tissus adipeux Download PDF

Info

Publication number
WO2016138132A1
WO2016138132A1 PCT/US2016/019361 US2016019361W WO2016138132A1 WO 2016138132 A1 WO2016138132 A1 WO 2016138132A1 US 2016019361 W US2016019361 W US 2016019361W WO 2016138132 A1 WO2016138132 A1 WO 2016138132A1
Authority
WO
WIPO (PCT)
Prior art keywords
dpp4
vat
diabetic
inflammation
mice
Prior art date
Application number
PCT/US2016/019361
Other languages
English (en)
Inventor
Ira A TABAS
Devram GHORPADE
Lale Ozcan
Original Assignee
The Trustees Of Columbia University In The City Of New York
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by The Trustees Of Columbia University In The City Of New York filed Critical The Trustees Of Columbia University In The City Of New York
Publication of WO2016138132A1 publication Critical patent/WO2016138132A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5067—Liver cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/948—Hydrolases (3) acting on peptide bonds (3.4)

Definitions

  • DIPEPTIDYL PEPTIDASE-IV (DPP4) INHIBITORS METHODS AND COMPOSITIONS FOR SUPPRESSING ADIPOSE TISSUE INFLAMMATION CROSS-REFERENCE TO RELATED APPLICATIONS
  • the invention relates generally to blocking dipeptidyl-peptidase 4 (DPP4) secretion/synthesis in hepatic cells, which in turn inhibits or blocks adipose tissue inflammation.
  • DPP4 dipeptidyl-peptidase 4
  • This process is involved in metabolism and type 2 diabetes, and will be useful for developing new DPP4 inhibitors and for treating or preventing conditions such as type 2 diabetes, metabolic syndrome, obesity, insulin resistance, as well as pre diabetes, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, diabetic dyslipidemia, fatty liver disease including non-alcoholic fatty liver disease (NASH), and atherosclerosis.
  • NASH non-alcoholic fatty liver disease
  • Dipeptidyl peptidase-IV (DPP-IV, DPP-4, or DPP4) is a serine protease that belongs to a group of post-proline/aianine cleaving amino-dipeptidases. DPP4 catalyzes the release of an N- terminal dipeptide only from proteins with N-terminal penultimate proline or alanine.
  • DPP4 The physiological role of DPP4 has not been established fully. It is believed to play an important role in neuropeptide metabolism, T-cell activation, gastric ulceration, functional dyspepsia, obesity, appetite regulation, impaired fasting glucose (IFG), and diabetes.
  • DPP4 has been implicated in the control of glucose metabolism because its substrates include the insulinotropic hormones, glucagon like peptide-1 (GLP-1) and gastric inhibitory peptide (G1P), which are inactivated by removal of their two N-terminal amino acids.
  • GLP-1 glucagon like peptide-1
  • G1P gastric inhibitory peptide
  • Diabetic dyslipidemia is characterized by multiple lipoprotein defects, including moderately high serum levels of cholesterol and triglycerides, small LDL particles, and low levels of HDL cholesterol.
  • the results of recent clinical trials reveal beneficial effects of cholesterol- lowering therapy in diabetic and nondiabetic patients, thus supporting increased emphasis on treatment of diabetic dyslipidemia. This need for intensive treatment of diabetic dyslipidemia was advocated by the National Cholesterol Education Program's Adult Treatment Panel III.
  • Obesity is a well-known risk factor for the development of many very common diseases such as atherosclerosis, hypertension and diabetes.
  • the incidence of obese people and thereby also these diseases is increasing throughout the entire industrialized world.
  • Even mild obesity increases the risk for premature death, diabetes, hypertension, atherosclerosis, gallbladder disease and certain types of cancer.
  • the prevalence of obesity has increased significantly in the past few decades. Because of the high prevalence of obesity and its health consequences, its prevention and treatment should be a high public health priority.
  • initial weight loss is not an optimal therapeutic goal. Rather, the problem is that most obese patients eventually regain their weight.
  • An effective means to establish and/or sustain weight loss is the major challenge in the treatment of obesity today.
  • Diabetes mellitus is a metabolic disorder characterized by recurrent or persistent hyperglycemia (high blood glucose) and other signs, as distinct from a single disease or condition. Glucose level abnormalities can result in serious long-term complications, which include cardiovascular disease, chronic renal failure, retinal damage, nerve damage (of several lands), microvascular damage and obesity.
  • Type 1 diabetes also known as Insulin Dependent Diabetes Mellitus (IDDM)
  • IDDM Insulin Dependent Diabetes Mellitus
  • Type-2 diabetes previously known as adult-onset diabetes, maturity-onset diabetes, or Non-Insulin Dependent Diabetes Mellitus (NIDDM)— is due to a combination of increased hepatic glucose output, defective insulin secretion, and insulin resistance or reduced insulin sensitivity (defective responsiveness of tissues to insulin).
  • microvascular disease due to damage of small blood vessels
  • macrovascular disease due to damage of the arteries.
  • microvascular disease include diabetic retinopathy, neuropathy and nephropathy
  • macrovascular disease examples include coronary artery disease, stroke, peripheral vascular disease, and diabetic myonecrosis.
  • Diabetic retinopathy characterized by the growth of weakened blood vessels in the retina as well as macular edema (swelling of the macula), can lead to severe vision loss or blindness. Retinal damage (from microangiopathy) makes it the most common cause, of blindness among non-elderly adults in the US.
  • Diabetic neuropathy is characterized by compromised nerve function in the lower extremities. When combined with damaged blood vessels, diabetic neuropathy can lead to diabetic foot. Other forms of diabetic neuropathy may present as mononeuritis or autonomic neuropathy.
  • Diabetic nephropathy is characterized by damage to the kidney, which can lead to chronic renal failure, eventually requiring dialysis. Diabetes mellitus is the most common cause of adult kidney failure worldwide.
  • a high glycemic diet i.e., a diet that consists of meals that give high postprandial blood sugar
  • a high glycemic diet i.e., a diet that consists of meals that give high postprandial
  • DPP4 inhibitors are widely used in humans with type 2 diabetes (T2D), with the major mechanism of action presumed to be the stabilization of GLP1, which promotes insulin secretion from pancreatic beta cells.
  • T2D type 2 diabetes
  • DPP4-I have other beneficial effects on metabolism and the mechanisms are not known. 1
  • circulating levels of DPP4 correlate with BMI and insulin resistance in humans.”
  • T2D type 2 diabetes
  • VAT visceral adipose tissue
  • VAT visceral adipose tissue
  • the present invention relates to a method for decreasing adipose tissue inflammation, comprising blocking DPP4 secretion from hepatocytes in the tissue of a subject in need thereof.
  • the blocking comprises administering an antagonist of DPP4.
  • the blocking comprises hepatic cell-specific genetic knockdown or knockout of DPP4.
  • the method further comprises inhibiting PAR2 binding to DPP4 in the tissue.
  • inhibiting PAR2 binding comprises contacting the tissue with an antagonist of PAR2.
  • the present invention relates to a method for decreasing adipose tissue inflammation comprising administering to a subject in need thereof, a compound that blocks DPP4 secretion from hepatocytes. In certain embodiments, the present invention relates to a method for preventing or alleviating obesity comprising administering to a subject in need thereof, a compound that blocks DPP4 secretion from hepatocytes.
  • the present invention relates to a method for preventing or alleviating metabolic syndrome comprising administering to a subject in need thereof, a compound that blocks DPP4 secretion from hepatocytes.
  • the subject is suffering from one or more conditions selected from the group consisting of: type 2 diabetes, metabolic syndrome, pre diabetes, obesity, insulin resistance, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, diabetic dyslipidemia, fatty liver disease including non-alcoholic fatty liver disease (NASH), and atherosclerosis.
  • NASH non-alcoholic fatty liver disease
  • the compound or blocking is prophylactically administered prior to symptoms normally associated with conditions selected from the group consisting of type 2 diabetes, metabolic syndrome, pre diabetes, obesity, insulin resistance, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, diabetic dyslipidemia, fatty liver disease including non-alcoholic fatty liver disease (NASH), and atherosclerosis.
  • diseases selected from the group consisting of type 2 diabetes, metabolic syndrome, pre diabetes, obesity, insulin resistance, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, diabetic dyslipidemia, fatty liver disease including non-alcoholic fatty liver disease (NASH), and atherosclerosis.
  • NASH non-alcoholic fatty liver disease
  • the method further improves insulin sensitivity. In additional embodiments, the method further suppresses VAT inflammation.
  • the present invention relates to a method for identifying a compound that blocks DPP4 secretion from hepatocytes comprising: a) incubating a hepatocyte culture with palmitate to induce DPP4 secretion, b) measuring secreted DPP4 in the induced hepatocyte culture, c) contacting and incubating the induced hepatocytes with a test compound, and d) measuring secreted DPP4 in the induced hepatocyte culture following the contacting and incubating step c), wherein a decrease in secreted DPP4 in the induced hepatocyte culture indicates a compound that blocks DPP4 secretion from induced hepatocytes, and wherein the decrease is a level similar to a reference basal DPP4 level observed in medium from non- induced hepatocytes.
  • the measuring is by ELISA.
  • FIGS. 1A-F illustrate that hepatocyte (HC)-specific CaMKII deletion lowers visceral adipose tissue (VAT) inflammation in diet-induced obese (DIO) mice, whereas hepatic and systemic inflammation are not affected.
  • DIO diet-induced obese mice
  • HC-CaMKII KO and WT mice were treated with adeno-associated viruses (AAV) containing either hepatocyte-specific TBG-Cre recombinase or the control vector (TBG-LacZ) to obtain HC-CaMKII KO and WT mice, respectively.
  • Fig. 1A shows representative images of VAT sections stained by immunohistochemistry with an antibody against the macrophage marker F4/80 after three weeks of AAV treatment.
  • lB-C are graphs showing shVAT and liver F4/80, Mcpl, and Tnf-a mRNA levels were assayed by RT-qPCR after three weeks of AAV treatment.
  • Fig. ID are graphs showing plasma 1L-6 and Tnf-a levels determined using a luminex assay system after three weeks of AAV treatment.
  • the 1HC data shown in Fig. IF used the Thirteen-week-old DIO Camklg ⁇ mice treated as in Fig. 1A.
  • FIGS. 2A-D show that the suppression of VAT inflammation in DIO HC-
  • FIG. 2A-B are graphs showing rsults from twelve-week-old DIO Camk2g m mice treated with TBG-Cre or TBG-LacZ to obtain HC-CaMKII KO and WT mice, respectively.
  • 5 hr fasting and fasted-refed blood glucose and plasma insulin were assayed after four weeks of AAV treatment.
  • FIG. 2C shows representative images of hematoxyiin-eosin (H&E) staining of VAT sections after four weeks of AAV treatment.
  • Figures 3A-C show that suppression of VAT inflammation by HC CaMKII deficiency is abrogated by restoring ATF4, whereas liver inflammation is not affected.
  • FIG. 3A shows images of VAT sections from twelve-week-old Camk2g x DIO mice treated with TBG-Cre or TBG-LacZ to obtain HC-CaMKII KO and WT mice, respectively. Two days later half of the TBG-Cre treated mice received adeno-ATF4, while the other half received adeno-LacZ control. Representative images of VAT sections stained by immunohistochemistry with an antibody against the macrophage marker F4/80 after four weeks of adenovirus treatment are shown. Figs.
  • FIGs 4A-C show that ATF4 expression in livers of lean mice is sufficient to induce VAT Mcpl.
  • Figs. 4A-B are western blots from 10- week-old WT lean mice treated with adeno-LacZ (Ad-LacZ) or adeno-ATF4 (Ad-ATF4) for two weeks and then liver and VAT extracts were assayed for ATF4 and ⁇ -actin by immunoblot.
  • FIG. 5A-G show that HC-derived VAT inflammatory activity can be detected in plasma using an ex-vivo adipose stromai-vascular-fraction (SVF) assay.
  • Fig. 5A are representative images of VAT sections from the VAT of lean and obese mice that were stained with H & E are shown.
  • Fig. 5B is a graph showing VAT Mcpl mRNA levels assayed by RT-qPCR.
  • FIG. 5C is a graph showing VAT stromal vascular fraction (SVF) cells obtained from a DIO mouse incubated with a medium containing 10% of lean or DIO mice plasma for 18 h and then Mcpl mRNA levels were assayed by RT-qPCR.
  • Fig. 5A are representative images of VAT sections from the VAT of lean and obese mice that were stained with H & E are shown.
  • Fig. 5B is a graph showing VAT Mcpl mRNA levels assayed by RT-qPCR.
  • FIG. 5D is a graph showing data from thirteen- week-old DIO Camk2g ft ' mice were treated with adeno- associated viruses (AAV) containing either hepalocyte-specific TBG-Cre recombinase or the control vector (TBG-LacZ) to obtain HC-CaMKII KO and WT mice, respectively.
  • Fig. 5E is a graph showing data from thirteen-week- old DIO Camklg ⁇ mice were treated as in Fig. 5D for three weeks.
  • Fig. 5F is a graph showing data from twelve-week-old Camk2g m DIO mice treated with TBG-Cre or TBG-LacZ to obtain HC-CaMKII KO and W r T mice, respectively. Two days later half of the TBG-Cre treated mice received adeno-ATF4, while the other half received adeno-LacZ control for four weeks.
  • VAT SVF cells from a DIO mouse were incubated with medium containing 10% of plasma obtained from indicated groups of mice and Mcpl levels were assayed by RT-qPCR.
  • Fig. 5G is a graph showing data from 10-week- old WT lean mice were treated with adeno-LacZ (Ad-LacZ) or adeno-ATF4 (Ad-.ATF4) for three weeks.
  • VAT SVF cells from a DIO mouse were incubated with medium containing 10% of pooled plasma obtained from indicated groups of mice and Mcpl levels were assayed by RT-qPCR (bars with different symbols are different from each other and from control, p ⁇ 0.05; mean ⁇ SEM).
  • FIG. 6A is a graph showing data from VAT SVF cells from a DIO mouse incubated with a medium containing 10 % control or heat-inactivated (HI) plasma from the indicated groups of mice for 18 h and then Mcpl mRNA levels were assayed by RT- qPCR.
  • Fig. 6B is a graph of UV protein chromatogram obtained after fractionation of lean and DIO mice plasma using size exclusion gel filtration fast flow chromatography is shown.
  • Fig. 6A is a graph showing data from VAT SVF cells from a DIO mouse incubated with a medium containing 10 % control or heat-inactivated (HI) plasma from the indicated groups of mice for 18 h and then Mcpl mRNA levels were assayed by RT- qPCR.
  • Fig. 6B is a graph of UV protein chromatogram obtained after fractionation of lean and DIO mice plasma using size exclusion gel filtration fast flow chromatography is shown.
  • FIG. 6C is a graph showing data from VAT SVF cells obtained from a DIO mouse were incubated with a medium containing 10 % of unfractionated plasma or plasma FPLC fractions corresponding to chromatogram peak in the 150 kDa-400 kDa range of protein separation from lean or DIO mice for 18 h and then Mcpl rnRNA levels were assayed by RT- qPCR.
  • Fig. 6D-F are graphs showing data from tsvelve-week-old Camk2g m DIO mice were treated with TBG-Cre or TBG-LacZ to obtain HC-CaMKII KO and WT mice, respectively.
  • VAT SVF cells obtained from a DIO mouse were incubated with medium containing 10% of unfractionated plasma or plasma FPLC fractions corresponding to chromatogram peak in the 150 kDa-400 kDa range of protein separation from WT, HC-CaMKll KO or ATF4 restored HC-CaMKII KO DIO mice for 18 h and then Mcpl mRNA levels were assayed by RT-qPCR.
  • 6G is a blot from obese DIO mouse plasma Mcpl inactive FPLC fractions (32, 34, 40, 41, 42, 46, 47, 48, 49, 58 or 61) or Mcpl active FPLC fractions (43, 44 or 45) were run on SDS PAGE reducing gel, stained with coomassie staining solution and destained using HPLC grade water. Two of Mcpl inactive fraction proteins (40 and 47) and one Mcpl active fraction proteins (44) were divided into 3 parts as marked and then subjected to in-gel trypsin digestion overnight. Tryptic-digested peptides were subjected to Fusion Tribrid Mass Spectrometer+Easy-nLC 1000 for global proteome analysis. Fig.
  • FIG. 6H is a graph showing DPP-4 levels detected after spectral analysis of indicated DIO mouse plasma fractions using LC-MS/MS.
  • Fig. 61 is a graph showing DPP- 4 activity measured in lean and DIO mice plasma using a f!uorometric DPP-4 activity assay.
  • Fig. 6J is a graph showing data from VAT SVF cells obtained from a DIO mouse incubated for 18 h with a medium containing 10% of DIO mice plasma with indicated doses of DPP-4 inhibitor (KR62436) after 1 hour pretreatment and Mcpl levels were assayed (bars with different symbols are different from each other and from control, p ⁇ 0.05; mean ⁇ SEM).
  • FIG. 7A-E are graphs showing that the CaMKII-ATF4 pathway regulates hepatic but not VAT DPP-4 expression and activity.
  • FIG. 7A-B are graphs showing results from twelve- week-old Camklg ⁇ DIO mice treated with TBG-Cre or TBG-LacZ to obtain HC-CaMKII KO and WT mice, respectively. Two days later half of the TBG-Cre treated mice received adeno-ATF4, while the other half received adeno-LacZ. Four weeks later, liver and VAT Dpp-4 mRNA levels were assayed by RT-qPCR. Fig.
  • FIG. 7C is a graph showing DPP- 4 activity measured in liver lysates obtained from VVT, HC-CaMKII KO or ATF4 restored HC-CaMKII KO DIO mice using a fluorometric DPP-4 activity assay.
  • Fig. 7D graph is similar to Fig. 7C except that VAT lysate was used. Fig.
  • 7E is a graph showing data from primary hepalocvtes from WT mice transduced with adeno-LacZ (Ad-LacZ) or adeno-ATF4 at an moi of 10 and 24 h later, Dpp4 mRNA levels were assayed by RT-qPCR (bars with different symbols are different from each other and from control, p ⁇ 0.05; mean ⁇ SEM).
  • FIG. 8A-E show that hepatic CaMKII-ATF4 pathway regulates plasma DPP- 4 activity.
  • Fig. 8A graph shows results from twelve-week-old Camk2gfl/fl DIO mice treated with TBG-Cre or TBG-LacZ to obtain HC-CaMKII KO and WT mice, respectively. Two days later half of the TBG-Cre treated mice received adeno-ATF4, while the other half received adeno-LacZ. Four weeks later, plasma from WT, CaMKII KO or ATF4 restored HC-CaMKII KO DIO mice were used to assay activity (Fig. 8A) of DPP-4 using a fluorometric DPP-4 activity assay .
  • Figs. 8B-D are graphs showing results from twelve-week- old Camk2gfl/fl DIO mice treated as in (Fig. 8A) for four weeks. DPP-4 activity was measured in plasma FPLC fractions corresponding to chromatogram peak in the 150 kDa-200 kDa range of protein separation from WT, HC-CaMKII KO or ATF4 restored HC-CaMKll KO DIO mice using a fluorometric DPP-4 activity assay.
  • Fig. 8E is a graph showing results from 10-week-old WT lean mice treated with adeno-LacZ (Ad-LacZ) or adeno-ATF4 (Ad- ATF4). Two weeks later, plasma was used to monitor DPP-4 activity (Fig. 8E) using a fluorometric DPP-4 activity assay (bars with different symbols are different from each other and from control, p ⁇ 0.05; mean + SEM).
  • ATMs adipose tissue macrophages
  • n containing 10 % of either lean or obese mice plasma
  • Figures 10A-D show that plasma from obese humans promotes VAT inflammation. Fig.
  • FIG. 10A is a graph showing data from obese mouse VAT SVF cells incubated for 18 h with 10% plasma obtained from lean or obese individuals and then Mcpl mRNA levels were assayed by RT-qPCR.
  • Fig. 10B is a graph showing data from obese mouse VAT SVF cells incubated with control plasma or heat inactivated (HI) plasma from lean or obese individuals for 18 h and then Mcpl mRNA levels were assayed by RT-qPCR.
  • Fig. IOC is a graph showing data from UV protein chromatogram obtained after fractionation of lean and obese human serum samples using size exclusion gel filtration fast flow chromatography. Fig.
  • 10D is a graph showing data from obese mouse VAT SVF cells incubated with unfractionated serum or FPLC fractions corresponding to chromatogram peak in the 150 kDa-400 kDa range of protein separation from lean or obese human serum and Mcpl mRNA levels were assayed by RT-qPCR (bars with different symbols are different from each other and from control, p ⁇ 0.05; mean ⁇ SEM).
  • FIGs 11A-B illustrate that HC specific deletion of ATF4 suppresses VAT inflammation.
  • Seventeen week old Atf4 fl il mice were treated with adeno-associated virus (AAV) containing either hepatocyte-specific TBG-Cre recombinase or the control vector (TBG-LacZ) to obtain HC-ATF4 KO and WT mice.
  • Fig. 11A shows representative images of VAT sections stained by immunohistochemistry with an antibody against the macrophage marker F4/80 after three weeks of AAV treatment.
  • CLS crown like structure
  • AAV adeno-associated virus
  • Fig. 12D is a graph showing 5h fasting plasma glucose levels in control and AAV8-ShDPP4 treated DIO mice.
  • Fig 12E shows the results obtained from glucose tolerance test (GTT) performed on control and AAV8-ShDPP4 treated DIO mice. Mice were fasted overnight and then injected with lg D-glucose/kg of mice and plasma glucose was measured at regular lime intervals as shown.
  • GTT glucose tolerance test
  • FIG. 12F is a graph showing plasma insulin levels obtained after 5h fasting of control or AAV8-ShDPP4 treated DIO mice.
  • Figures 13A-D show that liver specific deletion of DPP-4 lowers VAT inflammation. Seventeen week old DIO mice were injected with adeno-associated vims (AAV) containing either ShDPP4 or the control vector (ShLacZ).
  • AAV adeno-associated vims
  • FIG. 13A shows representative images of VAT sections stained by immunohistochemistry with an antibody against the macrophage marker F4/80 after four weeks of AAV8-LacZ or AAV8-ShDPP4 treatment.
  • Fig. 13B is a graph showing results obtained from quantification of crown like structure (CLS) macrophages in VAT sections.
  • FIG. 13D shows an immunoblol obtained from VAT tissues of either control or AAV8-ShDPP4 treated obese mice.
  • VAT tissue extracts were obtained after four weeks of adeno associated virus treatment and activation of Akt pathway was assayed after 4 min of portal vein injection of 0.7 IU/kg of insulin.
  • FIG. 14A-C show that obese pIasma-DPP-4 triggered inflammation in stromal vascular fraction (SVF) cells and M(j>s is reduced by PAR2 inhibition.
  • Fig 14C shows data obtained from obese VAT SVF cells. Obese plasma was incubated with either protein G beads conjugated anti-DPP4 antibody or control IgG for 2h on rotating shaker. Immunoprecipitated DPP-4 was then eluted using elution buffer.
  • BMDM bone marrow derived macrophage
  • FIG. 15A-C show that DPP4 needs to be complexed with one or more plasma proteins to cause VAT inflammation.
  • Fig. ISA is a graph showing data from obese VAT SVF cells, which were either treated with mouse rDPP-4 alone or in combination with obese mouse plasma for 4h and then Mcpl mRNA levels were assayed using RT-qPCR.
  • Fig. 15B shows results obtained from obese VAT SVF cells which were treated with either obese plasma, DPP-4 depleted obese plasma or combination of DPP-4 depleted obese plasma and mouse rDPP-4 or mouse rDPP4 alone for 4h and then Mcpl mRNA levels were assayed using RT-qPCR.
  • FIG. 16A graphically represents Dpp4 mRNA and summarizes strategy to design primers for determining spliced forms of Dpp4 mRNA.
  • the present invention relates to methods for decreasing adipose tissue inflammation comprising administering to a subject, a compound that blocks DPP4 secretion from hepatocytes.
  • Blocking DPP4 synthesis in hepatocytes has the downstream effect of blocking adipose tissue inflammation, which is beneficial in many aspects of metabolism including type 2 diabetes, as well as metabolic syndrome and conditions in which visceral adipose tissue (VAT) inflammation plays a role, including, pre diabetes, obesity, insulin resistance, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy, diabetic dysMpidemia, and potentially other metabolic diseases such as fatty liver disease including non-alcoholic fatty liver disease (NASH), and atherosclerosis.
  • NASH non-alcoholic fatty liver disease
  • the present invention relates to a method for preventing or alleviating obesity comprising administering to a subject, a compound that blocks DPP4 secretion from hepatocytes.
  • the present invention relates to a method for preventing or alleviating metabolic syndrome comprising administering to a subject, a compound that blocks DPP4 secretion from hepatocytes.
  • the method further improves insulin sensitivity.
  • the method further suppresses VAT inflammation.
  • liver DDP4 synthesis and circulating levels of DDP4 are suppressed in obese mice when hepatocyte ATF4 is silenced. This data is useful for elucidating the mechanisms of ATF4-mediated induction of DPP4.
  • the present data also show that DPP4 activates inflammatory pathways in macrophages and provides further elucidation of the mechanism involved in this process.
  • the present data also shows that blocking DPP4 secretion from hepatocytes in obese mice decreases adipose tissue inflammation and improves insulin sensitivity. While not being bound by theory, it is believed that the condition of obesity serves to stimulate DPP4 synthesis and secretion into the circulation, by activating the PERK-ATF4 EPv stress pathway in hepatocytes. In turn, DPP4 promotes adipose tissue inflammation by acting on adipose macrophages. Thus, in view of the present data, it is believed that blocking DDP4 secretion from hepatocytes in obese subjects will decrease adipose tissue inflammation and improve insulin sensitivity. Molecular biology
  • John Wiley and Sons, Inc. Hoboken, N.J.; Coligan et al. eds. (2005) Current Protocols in Immunology, John Wiley and Sons, Inc.: Hoboken, N.J.; Coico et al. eds. (2005) Current Protocols in Microbiology, John Wiley and Sons, Inc.: Hoboken, N.J.; Coligan et al. eds. (2005) Current Protocols in Protein Science, John Wiley and Sons, Inc.: Hoboken, N.J.; and Enna et al. eds. (2005) Current Protocols in Pharmacology, John Wiley and Sons, Inc.: Hoboken, N.J.
  • Treating” or “treatment” of a state, disorder or condition includes:
  • the benefit to a subject to be treated is either statistically significant or at least perceptible to the patient or to the physician.
  • prophylactically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.
  • Acceptable excipients, diluents, and carriers for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington: The Science and Practice of Pharmacy. Lippincott Williams & Wilkins (A. R. Gennaro edit. 2005). The choice of pharmaceutical excipient, diluent, and carrier can be selected with regard to the intended route of administration and standard pharmaceutical practice.
  • an “immune response” refers to the development in the host of a cellular and/or antibody-mediated immune response to a composition or vaccine of interest.
  • a response usually consists of the subject producing antibodies, B cells, helper T cells, suppressor T cells, regulatory T cells, and/or cytotoxic T cells directed specifically to an antigen or antigens included in the composition or vaccine of interest.
  • a “therapeutically effective amount” means the amount of a compound that, when administered to an animal for treating a state, disorder or condition, is sufficient to effect such treatment.
  • the “therapeutically effective amount” will vary depending on the compound, the disease and its severity and the age, weight, physical condition and responsiveness of the animal to be treated.
  • compositions of the invention may include a "therapeutically effective amount” or a “prophylactically effective amount” of a compound described herein.
  • a “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic result.
  • a therapeutically effective amount of an antibody or antibody portion may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the antibody or antibody portion to elicit a desired response in the individual.
  • a therapeutically effective amount is also one in which any toxic or detrimental effects of the compound are outweighed by the therapeutically beneficial effects.
  • a “prophylactically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.
  • the present invention provides a pharmaceutical composition or formulation comprising at least one active composition, or a pharmaceutically acceptable derivative thereof, in association with a pharmaceutically acceptable excipient, diluent and/or carrier.
  • a pharmaceutical composition or formulation comprising at least one active composition, or a pharmaceutically acceptable derivative thereof, in association with a pharmaceutically acceptable excipient, diluent and/or carrier.
  • the excipient, diluent and/or carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not deleterious to the recipient thereof.
  • the compositions of the invention can be formulated for administration in any convenient way for use in human or veterinary medicine. The invention therefore includes within its scope pharmaceutical compositions comprising a product of the present invention that is adapted for use in human or veterinary medicine.
  • the pharmaceutical composition is conveniently administered as an oral formulation.
  • Oral dosage forms are well known in the art and include tablets, caplets, gelcaps, capsules, and medical foods. Tablets, for example, can be made by well-known compression techniques using wet, dry, or fluidized bed granulation methods.
  • Such oral formulations may be presented for use in a conventional manner with the aid of one or more suitable excipients, diluents, and carriers.
  • Pharmaceutically acceptable excipients assist or make possible the formation of a dosage form for a bioactive material and include diluents, binding agents, lubricants, glidants, disinte grants, coloring agents, and other ingredients.
  • Preservatives, stabilizers, dyes and even flavoring agents may be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, ascorbic acid and esters of p-hydroxybenzoic acid.
  • Antioxidants and suspending agents may be also used.
  • An excipient is pharmaceutically acceptable if, in addition to performing its desired function, it is non-toxic, well tolerated upon ingestion, and does not interfere with absorption of bioactive materials.
  • Acceptable excipients, diluents, and carriers for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington: The Science and Practice of Pharmacy. Lippincott Williams & Wilkins (A. R. Gennaro edit. 2005). The choice of pharmaceutical excipient, diluent, and carrier can be selected with regard to the intended route of administration and standard pharmaceutical practice.
  • the phrase “pharmaceutically acceptable” refers to molecular entities and compositions that are "generally regarded as safe * ', e.g., that are physiologically tolerable and do not typically produce an allergic or similar untoward reaction, such as gastric upset, dizziness and the like, when administered to a human.
  • the term “'pharmaceutically acceptable” ' means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopoeia or other generally recognized pharmacopeias for use in animals, and more particularly in humans.
  • “Patient” or “subject” refers to mammals and includes human and veterinary subjects.
  • the dosage of the therapeutic formulation will vary widely, depending upon the nature of the disease, the patient's medical history, the frequency of administration, the manner of administration, the clearance of the agent from the host, and the like.
  • the initial dose may be larger, followed by smaller maintenance doses.
  • the dose may be administered as infrequently as weekly or biweekly, or fractionated into smaller doses and administered daily, semi- weekly, etc., to maintain an effective dosage level.
  • oral administration will require a higher dose than if administered intravenously.
  • topical administration will include application several times a day, as needed, for a number of days or weeks in order to provide an effective topical dose.
  • carrier refers to a diluent, adjuvant, excipient, or vehicle with which the compound is administered.
  • Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, olive oil, sesame oil and the like. Water or aqueous solution saline solutions and aqueous dextrose and glycerol solutions are preferably employed as carriers, particularly for injectable solutions.
  • the carrier can be a solid dosage form carrier, including but not limited to one or more of a binder (for compressed pills), a glidant, an encapsulating agent, a flavorant, and a colorant. Suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences” by E. W. Martin.
  • mice or DIO mice will be purchased from The Jackson Laboratory. Camk2g- fl mice were generated as described previously 8 . The mice will be maintained on a 12-h dark-light cycle with high-fat diet (Research diets) feeding for 17 weeks. AAV-TBG- Cr and adeno-ATF4 (0.5-3 x 10 plaque-forming units/mice) will be delivered by tail vein injection and experiments will be commenced after 5-7 days.
  • Real time PGR First strand cDNA synthesis will be made from RNA, and real time PGR will be performed using primers specific for Mcpl, F480, and Tnfa and SYBR green PGR reagent.
  • VAT will be fixed in 10 % buffered formalin solution, dehydrated with graded ethanol, cleared with xylene, and embedded in paraffin. Sections (5 mm) will be cut using a Leica RM2125 microtome and stained with hematoxylin and eosin. Images will be obtained using a Nikon Olympus DP25 microscope.
  • SVF assay SVF cells from obese VAT will be isolated as described 26 and cultured with 10 % plasma for 18 h. Total RNA will be isolated and assayed for Mcpl expression using real time PGR.
  • iTRAQ MS Protein samples (80 mg) will be reconstituted in dissolution buffer and trypsinized for 16 h at 37°C. Tryptic digests will be labeled with four different iTRAQ reagents according to the manufacturer's instructions (iTRAQ Reagents Multiplex kit; Applied Biosystems), pooled together, and subjected to strong cation exchange (SCX) fractionation. Eight fractions will be collected and subjected to MS analysis. The MS data will be analyzed using Proteome Discoverer (Thermo Fisher Scientific, Beta Version 1.2.0.208) and Sequest search algorithm, against the NCBI protein database. Relative quantitation of proteins will be carried out based on the relative intensities of reporter ions released during MS fragmentation of peptides.
  • Blocking this pathway specifically in HCs via HC-specific knockout of CaMKH causes a marked decrease in VAT inflammation, with >75% decrease of macrophages and crown-like structures (CLS), marked lowering of VAT inflammatory cytokines, and decreased monocyte uptake into VAT ⁇ See, Fig. 1). This effect was NOT simply due to the improvement in insulin sensitivity.
  • the responding cell type in the SVF assay is adipose tissue macrophages (ATMs), not pre-adipocytes.
  • ATMs adipose tissue macrophages
  • DPP4 activity and ELISA in the plasma of all of the above mouse models fit the pattern of ex-vivo SVF/macrophage-Mcpl -inducing activity in the plasma (which in turn corceiates with in vivo VAT inflammation as above), and it also fit the pattern of Dpp4 mRNA and DPP4 activity in the liver (See, Fig. 7A-E, See ako Figs 8-9). Moreover, Dpp4 mRNA is increased in primary HCs transduced with ATF4.
  • mice (1 ) AAV8-shAtf4; (2) Atf4-floxed mice (which we have) crossed with HC-specifie AAV8-TBG-cre mice; and controls (3) AAV8-LacZ and (4) AAV8-TBG-LacZ, respectively.
  • the assay endpoints include: (a) Atf4 and Dpp4 mRNA in liver; (b) SVF-Mcpl -inducing activity, DPP4 activity, and DPP4 ELISA in the plasma: and (c) VAT inflammation: macrophages, crown-like structures (CLS), inflammatory cytokines.
  • mice will be fed high-fat (DIO) diet with 60%' kcal from fat obtained from Research Diets (D12492), and they will be maintained on a 12 hr light-dark cycle.
  • the AAV8 vectors are delivered by tail vein injection at 2-3 X 10 11 genome copies/mouse.
  • the mice are monitored frequently for signs of distress by both the researchers in our lab and ICM staff. Euthanasia by CO2 asphyxia followed by cervical dislocation will be used at the end of the experiment and for mice in distress.
  • Luciferase reporter constructs driven by the Dpp4 promoter will be made, which will show that when the HCs are transduced with Atf4 (see data above), the luciferase reporter is induced.
  • the Dpp4 promoter does contain putative ATF4 binding sites, and ChIP assays based on this information will be performed to show that ATF4 occupancy of these sites in the various mouse models correlates with hepatic Dpp4 mRNA.
  • RNAseq will be used to determine whether any of the sequences immunoprecipitate with anti-ATF4 belong to the promoter of Dpp4 or other genomic regions that could regulate Dpp4 transcription.
  • ATF4 does not directly induce Dpp4 but rather induces a Dpp4 transcriptional activator or represses a Dpp4 repressor.
  • RNAseq will be conducted comparing obese mice +/- silenced HC ATF4 (above) and screening the results for transcription factors (or repressors) that match known regulators of Dpp4 or are known to bind cognate cis elements in the Dpp4 promoter.
  • One candidate, the ATF4 target CHOP has already been ruled out by recent experiments, but other indirect links are possible.
  • IL-6 and DPP4 are in the same pathway.
  • One possibility is that IL-6 functionally interacts with the PERK-ATF4 pathway to induce DPP4, i.e., a 2-hit model such that blocking either ATF4 or IL-6 blocks DPP4 induction.
  • DPP4 affects inflammatory signaling in adipose macrophages.
  • DPP4 promotes adipose tissue inflammation by activating inflammatory signaling pathways in adipose macrophages (ATMs).
  • ATMs adipose macrophages
  • the VAT inflammatory activity in obese plasma induces inflammation in macrophages in a manner that is blocked by a DPP4 inhibitor. This result could reflect a direct effect of DPP4 on macrophages or a secondary effect due to an action of DPP4 on another protein in the plasma, eg a substrate for its protease activity like GLP1 or G1P.
  • rDPP4 alone is not able to induce Mcpl mRNA, a marker of VAT inflammation in vivo and ex vivo, but DPP4 isolated from obese plasma by immunoprecipitation does induce Mcpl.
  • rDPP4 combined svifh DPP4 depleted obese plasma is able to induce VAT inflammation suggesting that DPP4 is required to interact with other plasma protein(s) to cause VAT inflammation. Further studies will be done to identify DPP4 partners which along with DPP4 act to induce VAT inflammation during obesity. For studies to date, KR-62436 from Sigma- Aldrich as the DPP4-1 has been used, but any DPP4-I relevant to clinical use and used in previous DIO mouse studies, such as des-fluoro-sitagliptin (DFS) can be used. Based on previous macrophage studies, the concentration will be 25 ⁇ .
  • DFS des-fluoro-sitagliptin
  • IP-DPP4 has a direct effect on macrophages
  • the inflammatory signaling hub that is activated will be identified by focusing on several likely candidates from the literature: NFkB, JNK, P38, ERK, PKC, and NLPR3 inflammasome, 3 ' 11 ' 3 with the possibility that more than one of these pathways is activated.
  • the phosphorylation state of P65, the 3 MAPKs, and PKC, and the levels of NLRP3 and IL- ⁇ will be assayed by Western blot in macrophages incubated with IP-DPP4 as above.
  • VAT inflammation Evaluating the role of the identified factors in VAT inflammation, including links to genetic polymorphisms and mutations could be further explored in obese human subjects. Determining how blocking hepatocyte DPP4 synthesis affects insulin sensitivity in obese mice. Blocking DDP4 secretion from hepatocytes in obese mice, by decreasing adipose tissue inflammation, will improve insulin sensitivity.
  • DFS-treated mice +/- restoration of only the VAT inflammatory pathway would be compared. Additional experiments directly comparing all VAT and metabolic parameters described above in 3 groups of DIO mice: (1) those with the DPP4 ⁇ macrophage inflammatory pathway inhibited as planned above; (2) DFS-treated; and (3) mouse model from (1) treated with DFS will be carried out. DFS will be added to the DIO diet a 1.1% by weight, based on studies in the literature. A significant but partial improvement in the mice with just the VAT inflammation pathway blocked is predicted, somewhat better improvement with the DFS, and a non- additive effect in the combined model. Assuming 3 groups of mice in triplicate, these experiments will require 90 mice based on the calculations above.
  • WT and mutant DIO mice will be assayed for the various markers of this pathway: plasma non-esterified fatty acids and glycerol, liver acetyl-CoA, and liver PC activity.
  • adipokines such as adiponectin that may link DPP4- induced VAT inflammation to insulin resistance in other tissues 1 ' 22 and so putative adipokines will be assayed in the present model. All in all, the eventual causation experiments should involve 2 groups of mice in triplicate and thus require 60 mice based on the calculations above.
  • DPP4 dipeptidyl peptidase 4

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Diabetes (AREA)
  • Hematology (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Obesity (AREA)
  • Biomedical Technology (AREA)
  • Immunology (AREA)
  • Endocrinology (AREA)
  • Emergency Medicine (AREA)
  • Urology & Nephrology (AREA)
  • Cell Biology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Microbiology (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Food Science & Technology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Epidemiology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne de manière générale le blocage de la sécrétion/synthèse de la dipeptidyl-peptidase 4 (DPP4) dans des cellules hépatiques, ce qui inhibe ou bloque à son tour l'inflammation des tissus adipeux. Ce procédé est impliqué dans le métabolisme et le diabète de type 2 et sera utile pour mettre au point de nouveaux inhibiteurs de DPP4 et pour traiter ou prévenir des affections telles que le diabète de type 2, le syndrome métabolique, l'obésité, la résistance à l'insuline ainsi que le prédiabète, la rétinopathie diabétique, la neuropathie diabétique, la néphropathie diabétique, la dyslipidémie diabétique, la stéatose hépatique, y compris la stéatose hépatique non alcoolique (NASH) et l'athérosclérose.
PCT/US2016/019361 2015-02-25 2016-02-24 Inhibiteurs de la dipeptidyl-peptidase iv (dpp4), procédés et compositions pour supprimer une inflammation des tissus adipeux WO2016138132A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201562120549P 2015-02-25 2015-02-25
US62/120,549 2015-02-25

Publications (1)

Publication Number Publication Date
WO2016138132A1 true WO2016138132A1 (fr) 2016-09-01

Family

ID=56789115

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2016/019361 WO2016138132A1 (fr) 2015-02-25 2016-02-24 Inhibiteurs de la dipeptidyl-peptidase iv (dpp4), procédés et compositions pour supprimer une inflammation des tissus adipeux

Country Status (1)

Country Link
WO (1) WO2016138132A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10030024B2 (en) 2013-09-25 2018-07-24 Vertex Pharmaceuticals Incorporated Imidazopyridazines useful as inhibitors of the PAR-2 signaling pathway
CN110240603A (zh) * 2018-03-09 2019-09-17 中国科学院上海药物研究所 一种噻吩[3,2-d]并嘧啶-4-酮类化合物的药物新用途
CN115109162A (zh) * 2022-06-28 2022-09-27 北京仁立竞合生物科技有限公司 含有间充质干细胞的药物组合物在减肥中的应用

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012159215A1 (fr) * 2011-05-24 2012-11-29 Polyvalor S.E.C. Compositions et procédés pour l'administration efficace et sûre d'arnsi à l'aide de nanocomplexes spécifiques à base de chitosane
US20130245091A1 (en) * 2010-02-24 2013-09-19 Arrowhead Madison Inc. Compositions for Targeted Delivery of siRNA
US20140228287A1 (en) * 2010-03-25 2014-08-14 Boehringer Ingelheim Vetmedica Gmbh 1-[(3-cyano-pyridin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-[3-(r)-amino-piperidin-1-yl]-axanthine for the treatment of a metabolic disorder of a predominantly carnivorous non-human animal
US20140315796A1 (en) * 2011-11-07 2014-10-23 The University Of Queensland Modulators of protease activated receptors

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130245091A1 (en) * 2010-02-24 2013-09-19 Arrowhead Madison Inc. Compositions for Targeted Delivery of siRNA
US20140228287A1 (en) * 2010-03-25 2014-08-14 Boehringer Ingelheim Vetmedica Gmbh 1-[(3-cyano-pyridin-2-yl)methyl]-3-methyl-7-(2-butyn-1-yl)-8-[3-(r)-amino-piperidin-1-yl]-axanthine for the treatment of a metabolic disorder of a predominantly carnivorous non-human animal
WO2012159215A1 (fr) * 2011-05-24 2012-11-29 Polyvalor S.E.C. Compositions et procédés pour l'administration efficace et sûre d'arnsi à l'aide de nanocomplexes spécifiques à base de chitosane
US20140315796A1 (en) * 2011-11-07 2014-10-23 The University Of Queensland Modulators of protease activated receptors

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MIYAZAKI ET AL.: "Increased hepatic expression of dipeptidyl peptidase-4 in non-alcoholic fatty liver disease and its association with insulin resistance and glucose metabolism", MOLECULAR MEDICINE REPORT, vol. 5, no. Iss. 3, 13 December 2011 (2011-12-13), pages 729 - 733 *
WRONKOWITZ ET AL.: "Soluble DPP4 induces inflammation and proliferation of human smooth muscle cells via protease-activated receptor 2", BIOCHIM BIOPHYS ACTA, vol. 1842, no. Iss. 9, 11 June 2014 (2014-06-11), pages 1613 - 1621, XP029014385, DOI: doi:10.1016/j.bbadis.2014.06.004 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10030024B2 (en) 2013-09-25 2018-07-24 Vertex Pharmaceuticals Incorporated Imidazopyridazines useful as inhibitors of the PAR-2 signaling pathway
CN110240603A (zh) * 2018-03-09 2019-09-17 中国科学院上海药物研究所 一种噻吩[3,2-d]并嘧啶-4-酮类化合物的药物新用途
CN115109162A (zh) * 2022-06-28 2022-09-27 北京仁立竞合生物科技有限公司 含有间充质干细胞的药物组合物在减肥中的应用
CN115109162B (zh) * 2022-06-28 2023-05-16 威海见生生物技术有限公司 含有间充质干细胞的药物组合物在减肥中的应用

Similar Documents

Publication Publication Date Title
Zhang et al. Gypenosides improve diabetic cardiomyopathy by inhibiting ROS‐mediated NLRP 3 inflammasome activation
Mukherjee et al. Nicotinamide adenine dinucleotide biosynthesis promotes liver regeneration
Birnbaum et al. Combined SGLT2 and DPP4 inhibition reduces the activation of the Nlrp3/ASC inflammasome and attenuates the development of diabetic nephropathy in mice with type 2 diabetes
Chang et al. Resveratrol inhibits NLRP3 inflammasome activation by preserving mitochondrial integrity and augmenting autophagy
Dobrian et al. Dipeptidyl peptidase IV inhibitor sitagliptin reduces local inflammation in adipose tissue and in pancreatic islets of obese mice
Fischer et al. Adalimumab prevents barrier dysfunction and antagonizes distinct effects of TNF-α on tight junction proteins and signaling pathways in intestinal epithelial cells
Wang et al. FAM3A activates PI3K p110α/Akt signaling to ameliorate hepatic gluconeogenesis and lipogenesis
Ye et al. Inflammation during obesity is not all bad: evidence from animal and human studies
Skeldon et al. Caspases and inflammasomes in metabolic inflammation
Chen et al. Effects of combining linagliptin treatment with BI-38335, a novel SGLT2 inhibitor, on pancreatic islet function and inflammation in db/db mice
Yu et al. Pattern Recognition Receptor‐Mediated Chronic Inflammation in the Development and Progression of Obesity‐Related Metabolic Diseases
Kim et al. Fimasartan, a novel angiotensin-receptor blocker, protects against renal inflammation and fibrosis in mice with unilateral ureteral obstruction: the possible role of Nrf2
Zhou et al. Toll-like receptor 5 signaling ameliorates liver fibrosis by inducing interferon β–modulated IL-1 receptor antagonist in mice
Liu et al. Effects and mechanisms of caffeine to improve immunological and metabolic abnormalities in diet-induced obese rats
Luo et al. Rutaecarpine derivative R3 attenuates atherosclerosis via inhibiting NLRP3 inflammasome‐related inflammation and modulating cholesterol transport
Galindo‐Hernández et al. Protein translation associated to PERK arm is a new target for regulation of metainflammation: A connection with hepatocyte cholesterol
Zilu et al. Effects of XIAP on high fat diet-induced hepatic steatosis: a mechanism involving NLRP3 inflammasome and oxidative stress
WO2016138132A1 (fr) Inhibiteurs de la dipeptidyl-peptidase iv (dpp4), procédés et compositions pour supprimer une inflammation des tissus adipeux
Cao et al. Sitagliptin reduces endothelial dysfunction and apoptosis induced by high-fat diet and palmitate in thoracic aortas and endothelial cells via ROS-ER stress-CHOP pathway
US11951081B2 (en) Metformin glycinate, pharmaceutical compositions comprising the same, and methods of using the same
Cherngwelling et al. Agomelatine protects against obesity-induced renal injury by inhibiting endoplasmic reticulum stress/apoptosis pathway in rats
Wang et al. Recent updates on targeting the molecular mediators of NAFLD
Zeng et al. Total Saponins from Nigella glandulifera Seeds Ameliorate Adjuvant‐Induced Rheumatoid Arthritis in Rats by Inhibition of an Inflammatory Response and Bone Erosion
Ahrens Antibodies in metabolic diseases
US20080279843A1 (en) Method For Improving Insulin Sensitivity By Administering an Inhibitor of Antitrypsin

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16756276

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16756276

Country of ref document: EP

Kind code of ref document: A1