WO2004084933A1 - Elevation of adenosine level by cytokine-induced expression of cd73 - Google Patents
Elevation of adenosine level by cytokine-induced expression of cd73 Download PDFInfo
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Definitions
- This invention relates to methods for inducing elevated adenosine levels in an individual and to treatment or prevention of diseases or disorders benefiting from an elevated adenosine level.
- the invention also concerns a method of up-regulating endothelial CD73 expression in an individual and to treatment or prevention of diseases or disorders benefiting from such CD73 up-regulation.
- lymphocytes and endothelial cells are a multi-step process.
- circulating cells use a very finely regulated set of adhesion molecules.
- Enhanced adhesion to endothelium and subsequent transmigration of re-circulating leukocytes through the endothelial lining of vessel wall into the tissue is characteristic for inflammation.
- the release of pro- and anti-inflammatory cytokines in a high extent takes place at sites of inflammation. Those cytokines are potent regulators of the expression of adhesion molecules.
- CD73 ecto-5'-nucleotidase
- ecto-5'-nucleotidase is a 70-kD glycosyl-phosphatidyl-inositol-anchored cell surface molecule with ecto-enzymatic activity. It is abundantly expressed on the vascular endothelium and at a low level on certain subpopulations of human lymphocytes. It is part of the purine salvage pathway by degrading nucleoside-5 '- monophosphates (AMP and IMP) into nucleotides like adenosine and inosine (1). Adenosine, a purine nucleoside product of the CD73 enzyme activity, binds to specific receptors on the cell surface.
- AMP and IMP nucleoside-5 '- monophosphates
- Adenosine is reported to have a role in many physiological and pathological events. So far four different subtypes of G protein- coupled adenosine receptors AIR, A2aR, A2bR and A3R have been cloned. Due to the diversity of the receptors and their abundant localization in different tissues, adenosine-adenosine receptor interaction leads to various physiological responses. Adenosine, by binding to Al and A2 receptors, regulates pathological consequences of inflammation by controlling leukocyte binding to endothelium and acts as an anti-inflammatory agent by binding to A2 and A3 receptors, through the inhibition of neutrophil degranulation (2). Adenosine also decreases eosinophil migration through activation of A3 receptor.
- Adenosine prevents cell damage during heart and central nervous ischemia (3-5). After hypoxia ecto-5'-nucleotidase activity increases due to phenomenon known as preconditioning. This results in release of large amounts of adenosine leading to increased resistance of cells to infarction for example in cardiac hypoxia.
- adenosine having an anti-inflammatory and cell protective effect, plays an important role in controlling the extent and consequences of inflammation, this work was designed to identify factors responsible for the regulation of CD73 expression as well as ecto-5'-nucleotidase mediated adenosine production.
- Adenosine as such could be administered to patients suffering from inflammatory conditions or conditions that untreated most likely would lead to tissue inflammation.
- a serious drawback by direct administration of adenosine is the rapid elimination of adenosine in vivo. Therefore, this work offers a new way to achieve elevated levels of adenosine over a prolonged time.
- the main object of the present invention is to provide a method for increase of the adenosine level in an individual and for maintaining the elevated adenosine level over a prolonged period of time and thereby prevent or treat inflammatory conditions or conditions that untreated would lead to tissue inflammation.
- Another object of this invention is to provide a method for up-regulating the expression of endothelial CD73 in an individual and for preventing or treating diseases or disorders benefiting from such CD73 up-regulation, that necessary not would require an elevated level of adenosine.
- this invention concerns a method of inducing an elevated level of adenosine in an individual by administering to said individual an effective amount of either i) recombinant protein CD73 or ii) a cytokine or another factor being capable of inducing endothelial CD73 expression, or a combination thereof.
- the invention concerns a method for prevention or treatment of a disease or disorder requiring or benefiting from the elevation of the adenosine level in an individual, by administering to said individual an effective amount of either i) recombinant protein CD73 or ii) a cytokine or another factor being capable of inducing endothelial CD73 expression, or a combination thereof.
- the invention concerns a method of up-regulating endothelial CD73 expression in an individual by administering to said individual an effective amount of a cytokine or another factor being capable of inducing endothelial CD73 expression.
- the invention concerns a method for prevention or treatment of a disease or disorder requiring or benefiting from up-regulating endothelial CD73 expression in an individual, by administering to said individual an effective amount of a cytokine or another factor being capable of inducing endothelial CD73 expression.
- Figure 1 is a graphic presentation showing that the induction of CD73 surface expression on HUVEC by IFN- ⁇ is both time and dose-dependent.
- HUVEC were exposed to lOOOU/ml IFN- ⁇ for indicated time periods, (b) HUVEC were cultured with different concentrations of IFN- ⁇ for 72 hours. Relative means of MFI ⁇ SEM of 3-9 experiments are shown. Control expression is the expression of CD73 without IFN- ⁇ at each time point. Background (the negative control staining) is subtracted. *P ⁇ 0.05; ** P ⁇ 0.01
- FIG. 2 is a microscopy photograph demonstrating that the induction of CD73 with IFN-alpha leads to increased expression rather than changes in its distribution.
- HUVEC were either grown in medium or induced with IFN-alpha for 72 hours and the cell surface expression of CD73 was detected with mAb 4G4 against CD73 and FITC conjugated anti-mouse IgG antibody, (a) On control HUVEC CD73 is expressed on the cell surface in a punctate like pattern, (b) After IFN- ⁇ induction CD73 is more intense, but surface distribution is similar as on control HUVEC. Staining with a negative control antibody 3G6 on control HUVEC (c) and after IFN- ⁇ induction (d). Original magnification 400X, Scale bar 10 ⁇ m.
- FIG 3 is a graphic presentation of the relative expression CD73 mRNA in HUVEC. HUVEC were incubated with or without IFN-alpha for 72 hours. Realtime RT-PCR analyses were performed with TaqMan. The figure represents the relative expression of CD73 mRNA between control and IFN-alpha treated cells. Normalization was performed using housekeeping gene GAPDH. The data present the means of three experiments made in duplicates ⁇ SEM, * P ⁇ 0.05.
- Figure 4 is a graph showing that the regulation of lymphoid CD73 expression differs from that of endothelial cells. U266B1 cells and PBL were grown in medium with or without 1000 U/ml IFN- ⁇ for different times.
- Figure 5 is a summary of semiquantitative analysis of immunohistochemical stainings of urine bladder samples from (Panel a) healthy and Panel b) tumour areas before and after IFN treatment is presented.
- Figure 6 shows microscopy photographs demonstrating the effect of IFN- ⁇ on expression of CD73 in bladder carcinoma
- (a) A bladder cancer specimen stained with anti-CD73 mAb 4G4 before IFN- ⁇ treatment. Some vessels are faintly or moderately expressing CD73.
- (b) The same tumour stained with anti-CD73 mAb 4G4 after IFN-alpha treatment. Vessels express CD73 moderately or abundantly
- d-f An example of a tumour (t) expressing CD73 before (d) and after (e) IFN-alpha treatment. Also in this case IFN-alphaincreased CD73 expression on endothelial cells,
- (f) Negative control staining Some vessels are marked with arrows.
- FIG. 7 shows that IFN- ⁇ increases cell-surface ecto-5'-nucleotidase activity.
- HUVEC (a) and PBL (b) were pre-treated for 48 hours without (open bars) or with 1000 U/ml IFN- ⁇ (closed bars).
- *P ⁇ 0.05 as compared to control cells
- the kinetic parameters V max and K m ) were calculated from the presented curves and summarized in the text.
- FIG. 8 shows the effect of IFN- ⁇ on permeability of HUVEC monolayers.
- Figure 9 are representative histograms depicting CD73 upregulation on human umbilical endothelial cells after incubation with IFN- ⁇ and - ⁇ .
- IFN- ⁇ increased the mean fluorescence intensity (MFI) from 26 to- 50, while during 48 hours of incubation IFN- ⁇ upregulated the MFI from 26 to 94.
- MFI mean fluorescence intensity
- X- axis is the fluorescence intensity in a logarithmic scale and y-axis is the relative number of cells.
- Figure 10 shows metabolic pathways regulating adenosine levels. The enzymatic reactions leading to the formation and degradation of adenosine are depicted.
- the amount of adenosine can be elevated by 1. upregulating/increasing amount of CD73; 2. providing more AMP; 3. inhibiting adenylate kinase, and 4. inhibiting adenosine deaminase, or combinations thereof.
- Figure 11 shows Hematoxylin-eosin stainings of formalin-fixed paraffin-embedded sections of lungs of the rats with multiorgan failure without treatment (A) or with IFN-beta and AMP treatment (B).
- This invention is based on screening of a range of potential mediators and finding that IFN- ⁇ , ⁇ and gamma are potent activators of CD73 expression.
- IFN-alpha was chosen for more detailed studies.
- IFN- ⁇ produced specific time and dose-dependent in vitro upregulation of CD73 expression on endothelium but not on PBL and even more importantly, it upregulated CD73 expression in tumour vessels of bladder carcinoma patients in vivo.
- Upregulation of endothelial CD73 after IFN- ⁇ induction was enzymatically functional producing adenosine from 5 '-AMP leading to enhanced barrier function in endothelial cells.
- a cell type specific difference in regulation of endothelial and lymphocyte CD73 was found.
- CD73 can be upregulated both in vitro and in vivo by cytokines, especially interferons.
- cytokines especially interferons.
- manipulation of its endogenous production via upregulation of CD73 may be a potential way to treat harmful inflammatory conditions such as e.g., reperfusion injuries in connection to myocardial infarction and stroke, organ transplantations and various tissue damages and traumas.
- the term “mediator” is meant to include any soluble factor that has an effect in the setting of inflammation.
- the term “inducer” is further meant to mean a factor that increases expression and enhances function of certain molecules.
- treatment shall be understood to include complete curing of a disease or disorder, as well as amelioration or alleviation of said disease or disorder.
- prevention shall be understood to include complete prevention, prophylaxis, as well as lowering the individual's risk of falling ill with said disease or disorder.
- This term shall also be understood to include preconditioning of tissue by elevating the adenosine level according to the method of this invention at a very early stage (e.g. before operations, before complete diagnosis at stroke and infarct patients) so as to prevent the tissue from damages.
- the term "individual” refers to a human or animal subject.
- an agent according to the present invention that is sufficient to bring about a desired therapeutical result, especially upon administration to an animal or human subject.
- Elevated level of adenosine shall be interpreted as an adenosine level that is at least 2 % higher, preferably at least 20 % higher, most preferably at least 30 % higher than the normal tissue level would be without the measures taken according to this invention.
- disease or disorder requiring or benefiting from an elevation of the adenosine level means that the prevention or treatment of said disease or disorder is facilitated by an elevated adenosine level.
- disease or disorder requiring or benefiting from up-regulating endothelial CD73 expression means that the prevention or treatment of said disease or disorder is facilitated by such up-regulation.
- inflammatory condition is meant to include any harmful and undesired inflammatory response in a tissue in an individual, wherein said inflammatory condition may result from an acute condition such as tissue trauma, a reperfusion injury resulting from myocardial infarction or stroke, organ transplantations or an other surgical operation, or from a chronic condition including allergic conditions, autoimmune diseases, and inflammatory diseases.
- Treatment of tissue traumas or reperfusion injuries shall in this invention particularly be understood as prevention of inflammatory conditions that most likely will follow if said traumas or reperfusion injuries are left untreated.
- an elevated level of adenosine in an individual can be induced by administering the recombinant CD73 protein, or by a cytokine or another factor capable of inducing endothelial CD73 expression or by a combination of both therapies, the use of a cytokine or another factor with similar capability in many cases would be preferable.
- administration of recombinant CD73 protein would be useful, in order to rapidly achieve an increased adenosine production, as an alternative or as an additional therapy.
- Suitable agents to be used in this invention include cytokines or other factors that directly or indirectly upregulate transcription of the CD73 gene.
- a suitable cytokine for use in this invention is typically an interferon or an interleukin, but also other agents may be used.
- the interferon may be alpha-, beta-, gamma-, omega-, or any other interferon and it can be any subtype of the aforementioned interferons. It is believed that particularly alpha- and beta- interferons are suitable for use in this invention.
- Any interleukin capable of inducing endothelial CD73 expression is also suitable for use in this invention. As examples of such interleukins can be mentioned IL-4, IL-10, IL-13 and IL-20.
- Typical diseases or disorders requiring or benefiting from elevation of the individual's adenosine levels are: tissue trauma; reperfusion injuries resulting from myocardial infarction or stroke, organ transplantations or other surgical operations; cancer or cancer metastasis; or inflammatory conditions resulting from the aforesaid traumas or reperfusion injuries or from chronic conditions including allergic conditions, autoimmune diseases, and inflammatory diseases.
- chronic conditions can be mentioned arthritis, allergic conditions such as asthma, inflammatory conditions such as inflammatory bowel disease or an inflammatory condition of the skin, psoriasis, Parkinson's disease, Alzheimer's disease, autoimmune diseases, type I or type II diabetes, atherosclerosis, multiple sclerosis, Crohn's disease, or rejection reactions due to organ transplantations.
- the administration of recombinant CD73 protein or a cytokine or both is combined with an administration of adenosine monophosphate (AMP) in order to safeguard the source for adenosine to be produced as result of the elevated CD73 level, obtained by elevated expression or by direct administering of the recombinant CD73 protein.
- AMP adenosine monophosphate
- the administration of recombinant CD73 protein or a cytokine or both is combined with an administration of an adenylate kinase inhibitor, which prevents AMP from conversion into adenosine diphosphate (ADP) or adenosine triphosphate (ATP).
- a combined administration of recombinant CD73 protein or a cytokine or both, with AMP and such an adenylate kinase inhibitor may be particularly preferred.
- the administration of recombinant CD73 protein or a cytokine or both is combined with an administration of an adenosine deaminase inhibitor which prevents the decomposition of adenosine.
- an adenosine deaminase inhibitor which prevents the decomposition of adenosine.
- This could also further be combined with administration of AMP and optionally also an adenylate kinase inhibitor which prevents AMP from conversion into adenosine diphosphate (ADP) or adenosine triphosphate (ATP).
- ADP adenosine diphosphate
- ATP adenosine triphosphate
- administering of recombinant CD73 protein or a cytokine or both is started as soon as a trauma patient or infarction or stroke patient is brought to care, optionally even if the final diagnosis is not fully clarified.
- the adenosine level can be increased as rapidly as possible.
- an adenylate kinase inhibitor and/or an adenosine deaminase inhibitor could be administrated in addition to the agents mentioned above.
- the therapeutically effective amount of the components according to this invention to be given to a patient in need of such treatment may depend upon a number of factors including, for example, the age and weight of the patient, the precise condition requiring treatment and its severity, and the route of administration. The precise amount will ultimately be at the discretion of the attending physician.
- practice of the present invention may involve any dose, combination with other therapeutically effective drugs, pharmaceutical formulation or delivery system for oral, topical, inhalation or parenteral administration.
- Amounts and regimens for the administration of the agents according to the present invention can be determined readily by those with ordinary skill in the art of treating inflammation-related disorders, such as reperfusion injuries, stroke, organ transplantation, traumas, cancer or cancer metastasis, or chronic inflammatory diseases.
- the cytokines or other factors may according to the present invention preferably be administered by infusion or by injection.
- Intravascular infusions are normally carried out using parenteral solutions contained within an infusion bag or bottle, and may be connected to different systems to control the rate of administration of the parenteral solution.
- the cytokines or other factors may according to the present invention alternatively be administered as an aerosol.
- Preferred formulations for infusion or injection may include carriers, such as human serum albumin, pharmaceutically acceptable salts, buffers, such as phosphates and or other pharmaceutically acceptable excipients.
- the active ingredient e.g. the cytokines or other factors may be provided in amounts ranging from e.g., 1-50 x 10 6 IU per ml.
- the formulation may preferably be provided as lyophilised powder in dosage form, to be prepared by the addition of water or other solutions suitable for injection prior to the administration.
- Cytokines such as interferons, or other factors with similar capability can be given to the patients suffering from or being at risk of getting inflammations.
- Those types of inflammatory conditions are for example ischemia reperfusion injuries during the stroke and myocardial infarction.
- organ transplantation and trauma are occasions often associated with major inflammatory components. Due to their unique characteristics different cytokines have their preferential disease targets: beta interferons are the most suitable interferons for ischemia reperfusion injuries in stroke and myocardial infarction, whereas alfa-interferons may not be the drug of choice in myocardial infarction.
- a suitable administration route would be infusion or injection.
- a suitable daily dose is in the range 0.1 to 5.0 mg kg body weight.
- the adenosine monophosphate may be administered e.g. subcutaneously, intramuscularly, intravenously or transdermally.
- a typical daily dose may be in the range 0.1 to 100 mg/kg body weight.
- an optionally used adenylate kinase inhibitor or an adenosine deaminase inhibitor may be, for example, administered subcutaneously, intramuscularly, intravenously or transdermally.
- a typical daily dose of such inhibitors may be in the range 0.1 to 100 mg/kg body weight.
- Human umbilical vein endothelial cells were isolated and cultured on gelatin-coated cell culture flasks in complete medium.
- Human peripheral blood lymphocytes (PBL) from healthy volunteers were isolated using Ficoll-Hypaque (Histopague-1077; Pharmacia, Uppsala, Sweden).
- PBL, U266B1 cell line, and the HEC endothelial cell line were cultured in RPMI 1640 medium containing 10% FCS, 4 mM L-glutamine, 100 U/ml penicillin, and 100 ⁇ g/ml streptomycin.
- Anti-CD73 mAb 4G4 (mouse IgGl), anti- ICAM-1 mAb 5C3 (IgGl) and mAb 3G6 (mouse IgGl) against chicken T-cells as a negative control antibody were used.
- ⁇ , ⁇ methyleneadenosine 5 '-diphosphate (AMPCP) and adenosine 5 '-monophosphate (5 '-AMP) were from Sigma (Sigma Chemical Co., St. Louis, MO). Inductions and immunofluorescence stainings. Details about inductions are indicated in Table 1. For every time-point, a control flask was incubated without inducers.
- Non treated cells (MFI, ⁇ CD73 - MFI, neg. co)
- lymphocytes were permeabilized before immunofluorescence stainings by incubating them for 2 min in acetone at -20 °C.
- RNA analysis HUVEC were grown to confluency in cell culture flasks and PBL were isolated prior to inductions using Ficoll-Hypague. 1 x 10 7 cells of both HUVEC and PBL were incubated with IFNalpha 1000 U/ml in culture medium. A similar number of cells was left untreated. RNA was isolated using UltraspecTM-II RNA Isolation System (Biotecx Laboratories, Inc., Houston, TX) according to the instructions of the manufacturer. 1-2 ⁇ g of total RNA was Dnase I treated (Amplification Grade, Gibco BRL, Life Technologies, Gathersburg, MD).
- cDNA was made by using Superscript II Reverse Transcriptase (Gibco BRL, Life Technologies) according to the manufacturer ' s instructions. Prior to real-time RT- PCR measurement, samples were treated with Rnase H (Gibco BRL, Life Technologies). Primers and probes for GAPDH housekeeping gene were used as internal controls.
- CD73 primers and probes were designed using Primer Express computer software (PE Biosystems, Foster City, CA). CD73 primers 5 'CTG GGA GCT TAC GAT TTT GCA3 ' and 5 'CCT CGC TGG TCT GCT CCA3 ' and CD73 probe 5 'CCA ACG ACG TGC ACA GCC GG3' were used (MedProbe, St.
- the real-time RT-PCR measurements were performed using TaqMan® Universal PCR Master Mix (Applied Biosystems, Branchburg, New Jersey) and ABI PRISM 7700 Sequence Detector (Applied Biosystems).
- the expression of the housekeeping gene GAPDH was used as a reference for normalization and the relative increase of CD73 mRNA expression between control and IFN- ⁇ treated cells was calculated.
- Ecto-5'-nucleotidase assay Ecto-5'-nucleotidase activity was assayed by thin layer chromatography (TLC) as described previously (19). Briefly, the standard enzyme assay contained in a final volume of 120 ⁇ l RPMI 1640, 4-6 x 10 4 detached HUVEC (or 1 x 10 5 lymphoid cells), 5 mmol/L ⁇ -glycerophosphate, and the indicated concentrations of 5'-AMP with tracer [ 2"3 H] AMP (specific activity 18.6 Ci/mmol; Amersham). Incubation times were chosen to ensure the linearity of the reaction with time, so that the amount of the converted AMP did not exceed 7-10% of the initially introduced substrate.
- TLC thin layer chromatography
- HUVEC Permeability assays. To evaluate barrier function of confluent monolayers, HUVEC were seeded (50 000 cells/insert) on Transwell insert polycarbonate filters (6.5-mm diameter, 0.4- ⁇ m pore size; Costar, Cambridge, MA). The filters were treated for 1- 2 h with fibronectin and air-dried before seeding endothelial cells. Typically, monolayers were studied 4-5 days post seeding. HUVEC were either induced with IFN- ⁇ (100 U/ml) for 72 hours before the studies of monolayer permeability or grown in medium without IFN- ⁇ .
- FITC-labeled dextran 500 ⁇ g/ml, mol wt 70 000.
- Endothelial monolayers were pre-treated with AMP (50 ⁇ M) for 15 min before the FITC-dextran transport was initiated.
- AMP a specific inhibitor of ecto-5'-nucleotidase
- AMPCP 100 ⁇ M
- AMPCP was added to the upper and lower chambers 30 min before the transport was initiated by adding FITC-labeled dextran.
- the inserts were removed from the bottom chamber (Visiplate, Perkin Elmer Life Sciences) at the time points 10 min, 20 min, 30 min, 40 min and 100 min and FITC- labelled dextran was measured directly from the bottom chambers in a fluorometer (TEC AN Ultra fluorescence reader, Tecan, Austria) using 485 and 535 nm as the excitation and emission wavelengths, respectively.
- TEC AN Ultra fluorescence reader Tecan, Austria
- IFN-alpha 1,5,10,50,100,200, 72 h 500 and 2000 U/ml 1000 u/ml 4,12,20,24,48, 60,72 and 96 h
- Immunofluorescence stainings followed by fluorescence microscopy revealed that IFN- ⁇ treatment does not induce any significant changes in the distribution or polarization of CD73 on HUVEC surface. Instead, CD73 is more intensely but similarly distributed on the cell surface ( Figure 2).
- tissue specimens from superficial epithelial bladder cancers were collected before and after IFN- ⁇ 2b treatment, stained and analysed.
- the bladder sample specimens were snap-frozen in liquid nitrogen and cut into 5 ⁇ m sections. Subsequently, sections were stained with anti-CD73 mAb 4G4 or 3G6 (negative control) as primary antibodies and peroxidase-conjugated rabbit anti- mouse IgG (DAKO A/S, Glostrup, Denmark) was used as a second stage antibody.
- the reaction was developed by adding 3,3 '-diaminobenzidine tetrahydrochloride (Poly sciences, Inc., Warrington, PA) in PBS. All incubations were 20 min with saturating mAb concentrations followed by two washes with PBS.
- the number of positive vessels/microscopic field (x200) was counted and intensity of the staining was semi-quantitatively evaluated. A combined score from 0 to 3 was given to each sample. Score 0 was assigned to samples with no positive blood vessels and score 3 to samples with staining equal to inflamed tonsil. Scores 1 and 2 were adjusted to cover the staining patterns in between. All samples were read blindly.
- Interferons produce a time and dose-dependent long-term upregulation of CD73 on endothelial cells but not on lymphocytes both at protein and RNA levels. Moreover, CD73 mediated production of adenosine is increased after IFN- ⁇ treatment on endothelial cells resulting in a decrease in the permeability of these cells.
- the multiorgan failure was induced to rats (weight: 250 g) by clamping the mesenteric artery for 30 minutes. Thereafter, the reperfusion time was two hours.
- the rats in the treatment group were injected sybcutaneously with 10 000 units of IFN-beta 18-20 hours before the clamping of the artery.
- the animals received 37.5 mg AMP in 3 ml saline as a continuous intravenous infusion. Rats with induced multiorgan failure but without treatment served as controls.
- the histology of the lungs which is one of the major target organs in this experimental model was analyzed.
- cytokines, and especially interferons are relevant in vivo regulators of CD73 in the endothelial-leukocyte microenvironment and thus have a fundamental role in controlling the extent of inflammation via CD73- dependent adenosine production.
- the methods of the present invention can be incorporated in the form of a variety of embodiments, only a few of which are disclosed herein. It will be apparent for the expert skilled in the field that other embodiments exist and do not depart from the spirit of the invention. Thus, the described embodiments are illustrative and should not be construed as restrictive.
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- Chemical Kinetics & Catalysis (AREA)
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- Heart & Thoracic Surgery (AREA)
- Molecular Biology (AREA)
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- Oncology (AREA)
- Transplantation (AREA)
- Pulmonology (AREA)
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- Physical Education & Sports Medicine (AREA)
- Urology & Nephrology (AREA)
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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Abstract
Description
Claims
Priority Applications (11)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE602004027797T DE602004027797D1 (en) | 2003-03-28 | 2004-03-19 | INCREASE OF THE ADENOSINE MIRROR BY CYTOKIN-INDUCED EXPRESSION OF CD73 |
SI200431498T SI1608400T1 (en) | 2003-03-28 | 2004-03-19 | Elevation of adenosine level by cytokine-induced expression of cd73 |
DK04721913.4T DK1608400T3 (en) | 2003-03-28 | 2004-03-19 | Elevation of adenosine level by cytokine-induced expression of CD73 |
US10/546,653 US7534423B2 (en) | 2003-03-28 | 2004-03-19 | Method for inducing an elevated level of adenosine in an individual |
AT04721913T ATE471720T1 (en) | 2003-03-28 | 2004-03-19 | INCREASING ADENOSINE LEVELS THROUGH CYTOKINE-INDUCED EXPRESSION OF CD73 |
JP2006505622A JP4809762B2 (en) | 2003-03-28 | 2004-03-19 | Increased adenosine levels by cytokine-induced expression of CD73 |
PL04721913T PL1608400T3 (en) | 2003-03-28 | 2004-03-19 | Elevation of adenosine level by cytokine-induced expression of cd73 |
EP04721913A EP1608400B1 (en) | 2003-03-28 | 2004-03-19 | Elevation of adenosine level by cytokine-induced expression of cd73 |
CA2519465A CA2519465C (en) | 2003-03-28 | 2004-03-19 | Elevation of adenosine level by cytokine-induced expression of cd73 |
US11/242,937 US7727521B2 (en) | 2003-03-28 | 2005-10-05 | Method of ameliorating multi-organ failure resulting from ischemic reperfusion injury |
US12/429,436 US20090269303A1 (en) | 2003-03-28 | 2009-04-24 | Method for treating or preventing conditions by elevation of the adenosine level in an individual |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
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FI20030467 | 2003-03-28 | ||
FI20030467A FI20030467A0 (en) | 2003-03-28 | 2003-03-28 | Procedure for treating inflammatory conditions |
US51542503P | 2003-10-30 | 2003-10-30 | |
US60/515,425 | 2003-10-30 |
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US10546653 A-371-Of-International | 2004-03-19 | ||
US11/242,937 Continuation-In-Part US7727521B2 (en) | 2003-03-28 | 2005-10-05 | Method of ameliorating multi-organ failure resulting from ischemic reperfusion injury |
US12/429,436 Division US20090269303A1 (en) | 2003-03-28 | 2009-04-24 | Method for treating or preventing conditions by elevation of the adenosine level in an individual |
Publications (1)
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WO2004084933A1 true WO2004084933A1 (en) | 2004-10-07 |
Family
ID=33099764
Family Applications (1)
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PCT/FI2004/000158 WO2004084933A1 (en) | 2003-03-28 | 2004-03-19 | Elevation of adenosine level by cytokine-induced expression of cd73 |
Country Status (9)
Country | Link |
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US (3) | US7534423B2 (en) |
EP (1) | EP1608400B1 (en) |
JP (1) | JP2011225607A (en) |
AT (1) | ATE471720T1 (en) |
CA (1) | CA2519465C (en) |
CY (1) | CY1111215T1 (en) |
DE (1) | DE602004027797D1 (en) |
PL (1) | PL1608400T3 (en) |
WO (1) | WO2004084933A1 (en) |
Cited By (16)
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WO2007042602A1 (en) | 2005-10-07 | 2007-04-19 | Faron Pharmaceuticals Oy | Method for treating or preventing ischemia reperfusion injury or multi-organ failure |
WO2009106368A1 (en) * | 2008-02-29 | 2009-09-03 | Gelato Corporation N.V. | Method for increasing the activity of the immune system of a mammal at risk of inflammatory diseases |
US7727521B2 (en) | 2003-03-28 | 2010-06-01 | Faron Pharmaceuticals Oy | Method of ameliorating multi-organ failure resulting from ischemic reperfusion injury |
EP2201376A1 (en) * | 2007-10-24 | 2010-06-30 | Faron Pharmaceuticals OY | A new biomarker for monitoring development of diseases and assessing the efficacy of therapies |
WO2011026132A2 (en) | 2009-08-31 | 2011-03-03 | 1/3Acamplimmune, Inc. | Methods and compositions for the inhibition of transplant rejection |
WO2012001647A2 (en) | 2010-06-30 | 2012-01-05 | Compugen Ltd. | Polypeptides and uses thereof as a drug for treatment of multiple sclerosis, rheumatoid arthritis and other autoimmune disorders |
WO2013040552A2 (en) | 2011-09-16 | 2013-03-21 | Georgia Health Sciences University | Methods of promoting immune tolerance |
WO2014100439A2 (en) | 2012-12-19 | 2014-06-26 | Amplimmune, Inc. | B7-h4 specific antibodies, and compositions and methods of use thereof |
WO2017041053A1 (en) | 2015-09-04 | 2017-03-09 | Yale University | Polymeric bile acid nanocompositions targeting the pancreas and colon |
WO2017155981A1 (en) | 2016-03-07 | 2017-09-14 | Massachusetts Institute Of Technology | Protein-chaperoned t-cell vaccines |
WO2017173453A1 (en) | 2016-04-01 | 2017-10-05 | The Brigham And Women's Hospital, Inc. | Stimuli-responsive nanoparticles for biomedical applications |
WO2018027039A1 (en) | 2016-08-03 | 2018-02-08 | Nextcure, Inc. | Compositions and methods for modulating lair signal transduction |
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US11957673B2 (en) | 2017-09-07 | 2024-04-16 | Augusta University Research Institute, Inc. | Specific AKT3 activator and uses thereof |
EP4360714A2 (en) | 2016-09-21 | 2024-05-01 | Nextcure, Inc. | Antibodies for siglec-15 and methods of use thereof |
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AU2002336650B2 (en) | 2001-10-25 | 2008-06-05 | Emory University | Catheter for modified perfusion |
US20070160645A1 (en) * | 2001-10-25 | 2007-07-12 | Jakob Vinten-Johansen | PostConditioning System And Method For The Reduction Of Ischemic-Reperfusion Injury In The Heart And Other Organs |
JP2008525468A (en) * | 2004-12-22 | 2008-07-17 | エモリー・ユニバーシティ | Treatment adjuvants that enhance postconditioning organ protection |
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US20090130092A1 (en) | 2007-10-30 | 2009-05-21 | Pinsky David J | Nucleotide phosphate dissipation as a treatment for vascular disorders |
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WO2013082458A1 (en) | 2011-12-02 | 2013-06-06 | The Regents Of The University Of California | Reperfusion protection solution and uses thereof |
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-
2004
- 2004-03-19 US US10/546,653 patent/US7534423B2/en active Active
- 2004-03-19 DE DE602004027797T patent/DE602004027797D1/en not_active Expired - Lifetime
- 2004-03-19 AT AT04721913T patent/ATE471720T1/en active
- 2004-03-19 PL PL04721913T patent/PL1608400T3/en unknown
- 2004-03-19 CA CA2519465A patent/CA2519465C/en not_active Expired - Lifetime
- 2004-03-19 EP EP04721913A patent/EP1608400B1/en not_active Expired - Lifetime
- 2004-03-19 WO PCT/FI2004/000158 patent/WO2004084933A1/en active Application Filing
-
2005
- 2005-10-05 US US11/242,937 patent/US7727521B2/en active Active
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2009
- 2009-04-24 US US12/429,436 patent/US20090269303A1/en not_active Abandoned
-
2010
- 2010-08-17 CY CY20101100758T patent/CY1111215T1/en unknown
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Also Published As
Publication number | Publication date |
---|---|
EP1608400B1 (en) | 2010-06-23 |
US20060198821A1 (en) | 2006-09-07 |
CY1111215T1 (en) | 2015-06-11 |
US7727521B2 (en) | 2010-06-01 |
US20090269303A1 (en) | 2009-10-29 |
PL1608400T3 (en) | 2010-11-30 |
ATE471720T1 (en) | 2010-07-15 |
JP2011225607A (en) | 2011-11-10 |
US7534423B2 (en) | 2009-05-19 |
CA2519465A1 (en) | 2004-10-07 |
DE602004027797D1 (en) | 2010-08-05 |
US20060034801A1 (en) | 2006-02-16 |
EP1608400A1 (en) | 2005-12-28 |
CA2519465C (en) | 2016-08-02 |
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