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HEK293/A2A/Gα15 Human/Recombinant ADORA2A ; A2ar

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Biology Department of WuXi AppTec (Shanghai) Co., Ltd. ADORA2A (Adenosine A2A Receptor) Cell Line Name: HEK293/A2A/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADORA2A ; A2aR; ADORA2; RDC8 Accession Number: NM_000675 G coupling Gs Functional Assay-Calcium Flux Assay ADORA2A NECA 120 Agonist NECA CPA 2+ 100 Measurement: Intracellular Ca 80 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40 Activity (%) Activity 20 Agonist effect EC50: 5.4 nM 0 -20 -2 0 2 4 6 Log [Compound] nM ADORA2A 120 Antagonist CGS-15943 CGS-15943 100 2+ ZM241385 Measurement: Intracellular Ca 80 Detection method: FLIPR Fluorimetry 60 40 Incubation 25℃ 20 Inhibition (%) Inhibition Stimulant NECA: 40 nM 0 Antagonist effect IC50: 0.22 nM -20 -4 -2 0 2 4 Log [Compound] nM ADORA2A (Adenosine A2A Receptor) Cellular Background: HEK293 Species/Source: Human/Recombinant Gene Synonyms: ADORA2A ; A2aR; ADORA2; RDC8 Accession Number: NM_000675 Radio ligand binding Assay ADORA2A Ligand [3H] CGS 21680 125 NECA Ligand concentration 6 nM 100 CPA ZM241385 Kd (nM) 47.5 nM 75 CGS15943 Non specific NECA (10 μM) 50 Incubation 120 min/RT Inhibition % 25 Detection method Scintillation counting 0 Reference: NECA (IC50:82.0 nM) -4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, China Email: [email protected]; www.wuxiapptec.com Page 1 of 44 Biology Department of WuXi AppTec (Shanghai) Co., Ltd. Alpha1A (Alpha-1-adrenergic receptor) Cell Line Name: HEK293/Alpha1A Species/Source: Human/Recombinant Gene Synonyms: ADRA1A; ADRA1C; ADRA1L1; ALPHA1AAR Accession Number: NM_000680 (isoform 1) G coupling Gq Functional Assay-Calcium Flux Assay Alpha1A 120 Agonist (-)Epinephrine (-)-Epinephrine (E4642) 100 A-61603 2+ Measurement: Intracellular Ca 80 60 Detection method: FLIPR Fluorimetry 40 (%) Activity Incubation 25℃ 20 0 Agonist effect EC50: 4.0 nM -2 -1 0 1 2 3 4 Log [Compound] nM Alpha1A 5-Methylurapidil Antagonist 5-Methylurapidil 120 WB-4104 Measurement: Intracellular Ca2+ 100 Prazosin 80 BMY7378 Detection method: FLIPR Fluorimetry 60 Incubation 25℃ 40 Inhibition (%) Inhibition 20 Stimulant (-)Epinephrine : 6.3 nM 0 -2 0 2 4 6 Antagonist effect IC50: 3.6 nM Log [Compound] nM Radio ligand binding Assay Ligand [3H]prazosin Alpha1A 125 WB4101 Ligand concentration 0.2 nM 100 Epinephrine Prazosin Kd (nM) 0.09 nM 75 Non specific epinephrine (0.1 mM) 50 % Inhibition % 25 Incubation 60 min/RT 0 Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference: WB 4101 (IC50:0.28 Log [Compound] nM nM) 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, China Email: [email protected]; www.wuxiapptec.com Page 2 of 44 Biology Department of WuXi AppTec (Shanghai) Co., Ltd. Alpha2A (Alpha-2-adrenergic receptor) Cell Line Name: HEK293/Alpha2A/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADRA2A; ADRA2; ADRAR; ZNF32; ADRA2R; ALPHA2AAR Accession Number: NM_000681 (isoform 1) G coupling Gi Functional Assay-Calcium Flux Assay Alpha2A UK 14304 Agonist UK14304 120 Epinephrine (E4250) 100 Measurement: Intracellular Ca2+ 80 Detection method: FLIPR Fluorimetry 60 ℃ 40 Incubation 25 (%) Activity 20 Agonist effect EC50: 0.14 nM 0 -20 -2 0 2 Log [Compound] nM Alpha2A 125 Antagonist Yohimbine hydrochloride Yohimbin 100 RX 821002 2+ Measurement: Intracellular Ca 75 Prazosin Detection method: FLIPR Fluorimetry 50 25 Incubation 25℃ (%) Inhibition 0 Stimulant UK14304 : 4.3 nM -25 -2 -1 0 1 2 3 4 Antagonist effect IC50: 32.6 nM Log [Compound] nM Radio ligand binding Assay Alpha2A Ligand [3H]RX 821002 125 Epinephrine Ligand concentration 0.5 nM 100 Prazosin Yohimbin 75 Kd (nM) 0.46 nM RX821002 Non specific epinephrine (0.1 mM) 50 UK14304 Incubation 60 min/RT Inhibition % 25 0 Detection method Scintillation counting -2 -1 0 1 2 3 4 5 Reference: Rx81002(IC50:2.3 nM) Log [Compound] nM 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, China Email: [email protected]; www.wuxiapptec.com Page 3 of 44 Biology Department of WuXi AppTec (Shanghai) Co., Ltd. Beta1 (Beta-1 Adrenergic Receptor) Cell Line Name: HEK293/Beta1/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADRB1; RHR; B1AR; ADRB1R; BETA1AR Accession Number: NM_000684 G coupling Gs Functional Assay-Calcium Flux Assay Beta1 (-)Epinephrine 120 Isoproterenol Agonist (-)Epinephrine 100 Salbutamol hemisulfate Measurement: Intracellular Ca2+ 80 60 Detection method: FLIPR Fluorimetry 40 Activity (%) Activity Incubation 25℃ 20 0 Agonist effect EC50: 2.72 nM -20 -2 -1 0 1 2 3 4 5 Log [Compound] nM Beta1 120 Antagonist Carvedilol Carvedilol 100 (± )-Propranolol hydrochloride Measurement: Intracellular Ca2+ 80 Atenolol 60 ICI 118 551 hydrochloride Detection method: FLIPR Fluorimetry 40 20 Inhibition (%) Inhibition Incubation 25℃ 0 -20 Stimulant (-)-Epinephrine : 20 nM -2 -1 0 1 2 3 4 Log [Compound] nM Antagonist effect IC50: 4.75 nM Radio ligand binding Assay Beta1 125 Ligand [3H](-)CGP 12177 Atenolol Ligand concentration 0.3 nM 100 Salbutamol Propranolol 75 Kd (nM) 0.04 nM ICI118551 50 Non specific propranolol (2 µM) Isoproternol % Inhibition % 25 Incubation 120 min/RT 0 Detection method Scintillation counting -4 -3 -2 -1 0 1 2 3 4 5 Reference: Atenolol (IC50:857 nM) Log [Compound] nM 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, China Email: [email protected]; www.wuxiapptec.com Page 4 of 44 Biology Department of WuXi AppTec (Shanghai) Co., Ltd. Beta2 (Beta-2 Adrenergic Receptor) Cell Line Name: HEK293/Beta2/Gα15 Species/Source: Human/Recombinant Gene Synonyms: ADRB2; BAR; B2AR; ADRBR; ADRB2R; BETA2AR Accession Number: NM_000024 G coupling Gs/Gi Functional Assay-Calcium Flux Assay Beta2 Adrenaline Agonist Adrenaline 120 Isoproterenol 100 Salbutamol hemisulfate 2+ Measurement: Intracellular Ca 80 Detection method: FLIPR Fluorimetry 60 40 Incubation 25℃ (%) Activity 20 Agonist effect EC50: 0.11 nM 0 -20 -4 -3 -2 -1 0 1 2 3 4 Log [Compound] nM Beta2 Carvedilol Antagonist Carvedilol 125 (± )-Propranolol hydrochloride ICI 118551 Measurement: Intracellular Ca2+ 100 75 Detection method: FLIPR Fluorimetry 50 Incubation 25℃ 25 Inhibition (%) Inhibition Stimulant Adrenaline: 0.43 nM 0 -25 Antagonist effect IC50: 7.07 nM -2 -1 0 1 2 3 Log [Compound] nM Radio ligand binding Assay Beta2 Ligand [3H](-)CGP 12177 125 Atenolol Ligand concentration 0.3 nM 100 Salbutamol Propranolol Kd (nM) 0.1 nM 75 ICI118551 Non specific propranolol (2 µM) 50 Isoproternol Incubation 120 min/RT Inhibition % 25 Detection method Scintillation counting 0 Reference: ICI118551(IC50:3.3 nM) -4 -3 -2 -1 0 1 2 3 4 5 Log [Compound] nM 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, China Email: [email protected]; www.wuxiapptec.com Page 5 of 44 Biology Department of WuXi AppTec (Shanghai) Co., Ltd. CB1 (Cannabinoid Type 1 Receptor) Cell Line Name: CHO/CB1/Gα15 Species/Source: Human/Recombinant Gene Synonyms: CNR1; CB1; CNR; CB-R; CB1A; CB1R; CANN6; CB1K5 Accession Number: NM_016083 (isoform1) G coupling Gi Functional Assay-Calcium Flux Assay CB1 WIN55212-2 Agonist WIN55212-2 120 CP55940 2+ 100 Anandamide Measurement: Intracellular Ca 80 Detection method: FLIPR Fluorimetry 60 40 Incubation 25℃ (%) Activity 20 Agonist effect EC50:352 nM 0 -20 -1 0 1 2 3 4 5 6 Log [Compound] nM CB1 Antagonist Rimonabant hydrochloride Rimonabant 120 AM281 2+ Measurement: Intracellular Ca 100 AM630 Detection method: FLIPR Fluorimetry 80 60 Incubation 25℃ 40 20 Inhibition (%) Inhibition Stimulant WIN55212-2: 4000 nM 0 -20 Antagonist effect IC50: 12.5 nM -1 0 1 2 3 4 5 6 log [Con.] nM CB1 (Cannabinoid Type 1 Receptor) Cellular Background: CHO-K1 Species/Source: Human/Recombinant Gene Synonyms: CNR1; CB1; CNR; CB-R; CB1A; CB1R; CANN6; CB1K5 Accession Number: X54937 Radio ligand binding Assay Ligand [3H]CP 55940 CB1 125 Ligand concentration 0.4 nM CP55940 100 WIN55212-2 AM281 Kd (nM) 3.7 nM 75 Anandamide Non specific WIN 55212-2 (5 µM) 50 Incubation 120 min/37℃ Inhibition % 25 0 Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference: CP 55940 (0.81 nM) Log [Compound] nM 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, China Email: [email protected]; www.wuxiapptec.com Page 6 of 44 Biology Department of WuXi AppTec (Shanghai) Co., Ltd. CB2 (Cannabinoid Type 2 Receptor) Cell Line Name: CHO/CB2/Gα15 Species/Source: Human/Recombinant Gene Synonyms: CNR2; CB2; CX5; CB-2 Accession Number: NM_001841 G coupling Gi Functional Assay-Calcium Flux Assay CB2 120 Agonist CP55940 CP55940 2+ 100 WIN55215-2 Measurement: Intracellular Ca 80 Anandamide Detection method: FLIPR Fluorimetry 60 40 ℃ Activity (%) Incubation 25 20 Agonist effect EC50: 199 nM 0 -1 0 1 2 3 4 5 Log [Compound] nM Antagonist AM630 CB2 120 2+ AM630 Measurement: Intracellular Ca 100 AM281 Detection method: FLIPR Fluorimetry 80 Rimonbant 60 Incubation 25℃ 40 20 Stimulant CP55940: 300 nM (%) Inhibition 0 Antagonist effect IC50: 563 nM -20 0 1 2 3 4 5 Log [Compound] nM Radio ligand binding Assay Ligand [3H]CP 55940 CB2 125 Ligand concentration 0.5 nM CP55940 100 WIN55212-2 Anandomide Kd (nM) 5.1 nM 75 AM630 Non specific WIN 55212-2 (5 µM) 50 AM281 % Inhibition % 25 Incubation 120 min/37℃ 0 Detection method Scintillation counting -3 -2 -1 0 1 2 3 4 5 Reference: WIN55212-2(IC50: Log [Compound] nM 2.2nM) 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, China Email: [email protected]; www.wuxiapptec.com Page 7 of 44 Biology Department of WuXi AppTec (Shanghai) Co., Ltd. CCKA (Cholecystokinin A Receptor) Cell Line Name: HEK293/CCKA Species/Source: Human/Recombinant Gene
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  • Acetylcholine Signaling System in Progression of Lung Cancers

    Acetylcholine Signaling System in Progression of Lung Cancers

    Pharmacology & Therapeutics 194 (2019) 222–254 Contents lists available at ScienceDirect Pharmacology & Therapeutics journal homepage: www.elsevier.com/locate/pharmthera Acetylcholine signaling system in progression of lung cancers Jamie R. Friedman a,1, Stephen D. Richbart a,1,JustinC.Merritta,KathleenC.Browna, Nicholas A. Nolan a, Austin T. Akers a, Jamie K. Lau b, Zachary R. Robateau a, Sarah L. Miles a,PiyaliDasguptaa,⁎ a Department of Biomedical Sciences, Joan C. Edwards School of Medicine, 1700 Third Avenue, Huntington, WV 25755 b Biology Department, Center for the Sciences, Box 6931, Radford University, Radford, Virginia 24142 article info abstract Available online 3 October 2018 The neurotransmitter acetylcholine (ACh) acts as an autocrine growth factor for human lung cancer. Several lines of evidence show that lung cancer cells express all of the proteins required for the uptake of choline (choline Keywords: transporter 1, choline transporter-like proteins) synthesis of ACh (choline acetyltransferase, carnitine acetyl- Lung cancer transferase), transport of ACh (vesicular acetylcholine transport, OCTs, OCTNs) and degradation of ACh (acetyl- Acetylcholine cholinesterase, butyrylcholinesterase). The released ACh binds back to nicotinic (nAChRs) and muscarinic Cholinergic receptors on lung cancer cells to accelerate their proliferation, migration and invasion. Out of all components Proliferation of the cholinergic pathway, the nAChR-signaling has been studied the most intensely. The reason for this trend Invasion Anti-cancer drugs is due to genome-wide data studies showing that nicotinic receptor subtypes are involved in lung cancer risk, the relationship between cigarette smoke and lung cancer risk as well as the rising popularity of electronic ciga- rettes considered by many as a “safe” alternative to smoking.
  • In Vivoactivation of a Mutantμ-Opioid Receptor by Naltrexone Produces A

    In Vivoactivation of a Mutantμ-Opioid Receptor by Naltrexone Produces A

    The Journal of Neuroscience, March 23, 2005 • 25(12):3229–3233 • 3229 Brief Communication In Vivo Activation of a Mutant ␮-Opioid Receptor by Naltrexone Produces a Potent Analgesic Effect But No Tolerance: Role of ␮-Receptor Activation and ␦-Receptor Blockade in Morphine Tolerance Sabita Roy, Xiaohong Guo, Jennifer Kelschenbach, Yuxiu Liu, and Horace H. Loh Department of Pharmacology, University of Minnesota, Minneapolis, Minnesota 55455 Opioid analgesics are the standard therapeutic agents for the treatment of pain, but their prolonged use is limited because of the development of tolerance and dependence. Recently, we reported the development of a ␮-opioid receptor knock-in (KI) mouse in which the ␮-opioid receptor was replaced by a mutant receptor (S196A) using a homologous recombination gene-targeting strategy. In these animals, the opioid antagonist naltrexone elicited antinociceptive effects similar to those of partial agonists acting in wild-type (WT) mice; however, development of tolerance and physical dependence were greatly reduced. In this study, we test the hypothesis that the failure of naltrexone to produce tolerance in these KI mice is attributable to its simultaneous inhibition of ␦-opioid receptors and activation of ␮-opioid receptors. Simultaneous implantation of a morphine pellet and continuous infusion of the ␦-opioid receptor antagonist naltrindole prevented tolerance development to morphine in both WT and KI animals. Moreover, administration of SNC-80 [(ϩ)-4-[(␣R)-␣-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-methoxybenzyl]-N,N-diethylbenzamide], a ␦ agonist, in the naltrexone- pelleted KI animals resulted in a dose-dependent induction in tolerance development to both morphine- and naltrexone-induced anal- gesia. We conclude that although simultaneous activation of both ␮- and ␦-opioid receptors results in tolerance development, ␮-opioid receptor activation in conjunction with ␦-opioid receptor blockade significantly attenuates the development of tolerance.
  • Intravta Deltorphin, but Not DPDPE, Induces Place Preference in Ethanoldrinking Rats

    Intravta Deltorphin, but Not DPDPE, Induces Place Preference in Ethanoldrinking Rats

    ALCOHOLISM:CLINICAL AND EXPERIMENTAL RESEARCH Vol. 38, No. 1 January 2014 Intra-VTA Deltorphin, But Not DPDPE, Induces Place Preference in Ethanol-Drinking Rats: Distinct DOR-1 and DOR-2 Mechanisms Control Ethanol Consumption and Reward Jennifer M. Mitchell, Elyssa B. Margolis, Allison R. Coker, Daicia C. Allen, and Howard L. Fields Background: While there is a growing body of evidence that the delta opioid receptor (DOR) modu- lates ethanol (EtOH) consumption, development of DOR-based medications is limited in part because there are 2 pharmacologically distinct DOR subtypes (DOR-1 and DOR-2) that can have opposing actions on behavior. Methods: We studied the behavioral influence of the DOR-1-selective agonist [D-Pen2,D-Pen5]- Enkephalin (DPDPE) and the DOR-2-selective agonist deltorphin microinjected into the ventral tegmental area (VTA) on EtOH consumption and conditioned place preference (CPP) and the physio- logical effects of these 2 DOR agonists on GABAergic synaptic transmission in VTA-containing brain slices from Lewis rats. Results: Neither deltorphin nor DPDPE induced a significant place preference in EtOH-na€ıve Lewis rats. However, deltorphin (but not DPDPE) induced a significant CPP in EtOH-drinking rats. In con- trast to the previous finding that intra-VTA DOR-1 activity inhibits EtOH consumption and that this inhibition correlates with a DPDPE-induced inhibition of GABA release, here we found no effect of DOR-2 activity on EtOH consumption nor was there a correlation between level of drinking and deltorphin-induced change in GABAergic synaptic transmission. Conclusions: These data indicate that the therapeutic potential of DOR agonists for alcohol abuse is through a selective action at the DOR-1 form of the receptor.