Adenosine receptor activation reagent and the uses of thereof

Abstract

The present invention relates to application of N(6)-(2-hydroxyethyl)-adenosine (HEA) and its derivatives as an adenosine A1 receptor agonist in preparation of drug or food, the HEA and its derivatives are used in treatment of diseases relating to adenosine receptor regulator, such as insomnia, pain, convulsion, apoplexia, Parkinson's disease, opioid drug addiction and kidney ischemia reperfusion injury etc. The present invention provides a new method for treatment of the diseases relating to nervous system and kidney.

Description

RELATED APPLICATIONS

[0001] The present application claims the priority from China Patent Application No. 201410360752.9 filed on Jul. 27, 2014, and China Patent Application No. 201510226745.4, filed on May 6, 2015. The entireties of these applications including all tables, diagrams and claims are incorporated hereby as reference of the present invention.TECHNICAL FIELD

[0002] The present invention relates to a new adenosine A1 receptor agonist, particularly, relates to N(6)-(2-hydroxyethyl)-adenosine (HEA) and its derivatives as adenosine A1 receptor agonist; and its new application as adenosine A1 receptor agonist in preparation of drug or food for prevention and treatment of the disease relating to adenosine receptor regulator.

[0003] Incorporated by reference herein in its entirety is the Sequence Listing entitled “sequence Listing.txt”, created Jul. 7, 2015, size of 2 kilobytes.BACKGROUND OF THE INVENTION

[0004] Cordyceps cicadae is a Cordyceps, in which the body is full of mycelium an...

Examples

example 1

HEA is a Selective Agonist of A1 Receptor

[0081]Preparation of Receptor Protein

[0082]Using Wistar rat, head was cut and the brain was removed, and cerebral cortex and striatum were separated, respectively weighed, added into 10 fold volume of ice-cooled Tris-HCL buffer solution (50 mM, PH7.5) according to 1:10, the tissue is homogenized, after the suspension was centrifuged the supernatant was discarded, the above-described solution wash was repeated for 3 times, centrifuged again and the supernatant was discarded, the precipitate was again mixed in 50 mM Tris-HCL buffer solution, the protein concentration in rat cerebral cortex determined by Coomassie Brilliant Blue Method (Bradford method) was 0.8 mg / ml, the protein content in rat striatum brain tissue homogenate was 1.3 mg / ml. After subpackage stored at −80□ for subsequent use (Li M, Kang R X, Shi J Liu G T, Zhang J J, 2013. Anticonvulsant Activity of B2, an Adenosine Analog, on Chemical Convulsant-Induced Seizures, PLoS One June ...

example 2

Application of HEA in Anti-Convulsion

[0090]2.1 Animal Model and Administration Method

[0091]Male ICR mouse, 18˜22 g; purchased from Animal Experiment Center of Wenzhou Medical University. Before experiment, animals were adapted to environment for at least 5 days. Kept at 25□, food and water were freely fed. Based on the weight, the healthy male ICR mice were randomly divided into control group (1% DMSO, ip), model group, CCPA group (0.1 mg·kg−1, ip), HEA group (15 mg / kg, 40 mg / kg, 60 mg / kg), DPCPX group (2 mg·kg−1, ip), ZM241385 group (1 mg·kg−1, 5 mg·kg−1, ip), DPCPX+HEA (2 mg·kg−1+40 mg / kg, ip) group and ZM241385+HEA (1 mg·kg−1+40 mg / kg, 5 mg·kg−1+40 mg / kg ip) group. Wherein, 10 min before administration, the adenosine A1R receptor antagonist DPCPX (or A2R receptor antagonist ZM241385) were peritoneal injected, after 15 min of administration pentylenetetrazole (100 mg·kg−1, ip) was given to induce mouse convulsion; in alone antagonist group, after 5 min of the antagonist pentylenet...

example 3

Application of the HEA in Cerebral Ischemia

[0096]3.1 Preparation of Tested Sample

[0097]The dried Cordyceps cicadae was precisely weighed as raw material, and extracted by using 50% ethanol as solvent, 2 h / time, filtered, the filtrate were combined, to prepare a solution containing sample with a effective dose of 1500 mg / kg for subsequent use.

[0098]3.2 Animal Model and Administration Method

[0099]Using rat middle cerebral artery distal obstruction cerebral ischemia model. (1) according to rat weight, 10% chloral hydrate (3 ml / kg) was peritoneally injected to anaesthetize. (2) the rat lie on right side and fixed, 1 cm skin incision was cut at connection of inner canthus with external canal, fascia was isolated, muscle tissue, the skull was exposed; (3) a small amount of normal saline was sucked with cotton ball to wipe the skull to ac clear view; (4) fascia was isolated under operating microscope, and the skull was exposed, a hole with diameter of 2 mm circular was drilled at a sute ⅓ ...

RELATED APPLICATIONS

[0001] The present application claims the priority from China Patent Application No. 201410360752.9 filed on Jul. 27, 2014, and China Patent Application No. 201510226745.4, filed on May 6, 2015. The entireties of these applications including all tables, diagrams and claims are incorporated hereby as reference of the present invention.TECHNICAL FIELD

[0002] The present invention relates to a new adenosine A1 receptor agonist, particularly, relates to N(6)-(2-hydroxyethyl)-adenosine (HEA) and its derivatives as adenosine A1 receptor agonist; and its new application as adenosine A1 receptor agonist in preparation of drug or food for prevention and treatment of the disease relating to adenosine receptor regulator.

[0003] Incorporated by reference herein in its entirety is the Sequence Listing entitled “sequence Listing.txt”, created Jul. 7, 2015, size of 2 kilobytes.BACKGROUND OF THE INVENTION

[0004] Cordyceps cicadae is a Cordyceps, in which the body is full of mycelium an...

Examples

example 1

HEA is a Selective Agonist of A1 Receptor

[0081]Preparation of Receptor Protein

[0082]Using Wistar rat, head was cut and the brain was removed, and cerebral cortex and striatum were separated, respectively weighed, added into 10 fold volume of ice-cooled Tris-HCL buffer solution (50 mM, PH7.5) according to 1:10, the tissue is homogenized, after the suspension was centrifuged the supernatant was discarded, the above-described solution wash was repeated for 3 times, centrifuged again and the supernatant was discarded, the precipitate was again mixed in 50 mM Tris-HCL buffer solution, the protein concentration in rat cerebral cortex determined by Coomassie Brilliant Blue Method (Bradford method) was 0.8 mg / ml, the protein content in rat striatum brain tissue homogenate was 1.3 mg / ml. After subpackage stored at −80□ for subsequent use (Li M, Kang R X, Shi J Liu G T, Zhang J J, 2013. Anticonvulsant Activity of B2, an Adenosine Analog, on Chemical Convulsant-Induced Seizures, PLoS One June ...

example 2

Application of HEA in Anti-Convulsion

[0090]2.1 Animal Model and Administration Method

[0091]Male ICR mouse, 18˜22 g; purchased from Animal Experiment Center of Wenzhou Medical University. Before experiment, animals were adapted to environment for at least 5 days. Kept at 25□, food and water were freely fed. Based on the weight, the healthy male ICR mice were randomly divided into control group (1% DMSO, ip), model group, CCPA group (0.1 mg·kg−1, ip), HEA group (15 mg / kg, 40 mg / kg, 60 mg / kg), DPCPX group (2 mg·kg−1, ip), ZM241385 group (1 mg·kg−1, 5 mg·kg−1, ip), DPCPX+HEA (2 mg·kg−1+40 mg / kg, ip) group and ZM241385+HEA (1 mg·kg−1+40 mg / kg, 5 mg·kg−1+40 mg / kg ip) group. Wherein, 10 min before administration, the adenosine A1R receptor antagonist DPCPX (or A2R receptor antagonist ZM241385) were peritoneal injected, after 15 min of administration pentylenetetrazole (100 mg·kg−1, ip) was given to induce mouse convulsion; in alone antagonist group, after 5 min of the antagonist pentylenet...

example 3

Application of the HEA in Cerebral Ischemia

[0096]3.1 Preparation of Tested Sample

[0097]The dried Cordyceps cicadae was precisely weighed as raw material, and extracted by using 50% ethanol as solvent, 2 h / time, filtered, the filtrate were combined, to prepare a solution containing sample with a effective dose of 1500 mg / kg for subsequent use.

[0098]3.2 Animal Model and Administration Method

[0099]Using rat middle cerebral artery distal obstruction cerebral ischemia model. (1) according to rat weight, 10% chloral hydrate (3 ml / kg) was peritoneally injected to anaesthetize. (2) the rat lie on right side and fixed, 1 cm skin incision was cut at connection of inner canthus with external canal, fascia was isolated, muscle tissue, the skull was exposed; (3) a small amount of normal saline was sucked with cotton ball to wipe the skull to ac clear view; (4) fascia was isolated under operating microscope, and the skull was exposed, a hole with diameter of 2 mm circular was drilled at a sute ⅓ ...

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