human CB2 receptor ( Ki = 45.5 nM ); human CB1 receptor ( Ki = 48.3 nM ); rat CB1 receptor ( Ki = 55.4 nM )

3-[2-Cyano-3-(trifluoromethyl)phenoxy]phenyl-4,4,4-trifluoro-1-butanesulfonate (BAY 59-3074) is a novel, selective cannabinoid CB1/CB2 receptor ligand (Ki = 55.4, 48.3, and 45.5 nM at rat and human cannabinoid CB1 and human CB2 receptors, respectively), with partial agonist properties at these receptors in guanosine 5-[γ35S]-thiophosphate triethyl-ammonium salt ([35S]GTPγS) binding assays[1].

BAY 59-3074 (oral administration; 0.3–3 mg/kg; daily; for 2 weeks; male Wistar rats) Rat models of chronic neuropathic and inflammatory pain show improved antihyperalgesic and antiallodynic effects against thermal or mechanical stimuli with treatment.

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BAY 59-3074 (0.3-3 mg/kg, p.o.) induced antihyperalgesic and antiallodynic effects against thermal or mechanical stimuli in rat models of chronic neuropathic (chronic constriction injury, spared nerve injury, tibial nerve injury, and spinal nerve ligation models) and inflammatory pain (carrageenan and complete Freund's adjuvant models). Antiallodynic efficacy of BAY 59-3074 (1 mg/kg, p.o.) in the spared nerve injury model was maintained after 2 weeks of daily administration. However, tolerance developed rapidly (within 5 days) for cannabinoid-related side effects, which occur at doses above 1 mg/kg (e.g., hypothermia). Uptitration from 1 to 32 mg/kg p.o. (doubling of daily dose every 4th day) prevented the occurrence of such side effects, whereas antihyperalgesic and antiallodynic efficacy was maintained/increased. No withdrawal symptoms were seen after abrupt withdrawal following 14 daily applications of 1 to 10 mg/kg p.o. It is concluded that BAY 59-3074 may offer a valuable therapeutic approach to treat diverse chronic pain conditions.[1]

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BAY 59-3074 [3-[2-cyano-3-(trifluoromethyl)phenoxy]phenyl-4,4,4-trifluoro-1-butane-sulfonate] is a structurally novel cannabinoid CB1/CB2 receptor partial agonist with analgesic properties. The present study was performed to confirm its receptor binding profile in a highly sensitive in vivo assay. Rats (n=10) learned to discriminate BAY 59-3074 (0.5 mg/kg, p.o., t-1 h) from vehicle in a fixed-ratio: 10, food-reinforced two-lever procedure after a median number of 28 training sessions. BAY 59-3074 generalized dose-dependently (ED(50): 0.081 mg/kg, p.o.) and the cue was detectable between 0.25 and 4 h after administration. The selective cannabinoid CB1 receptor antagonist SR 141716A [N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride] blocked the discriminative effects of BAY 59-3074 (ID50: 1.79 mg/kg, i.p.). Complete generalization was also obtained after i.p. administration of BAY 59-3074 (ED50 value: 0.41 mg/kg), and the reference cannabinoids BAY 38-7271 [(-)-(R)-3-(2-hydroxymethylindanyl-4-oxy)phenyl-4,4,4-trifluoro-1-butanesulfonate, 0.011 mg/kg], CP 55,940 [(-)-cis-3-[2-hydroxy-4(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxy-propyl)cyclohexanol, 0.013 mg/kg], HU-210 [(-)-11-OH-Delta8-tetrahydrocannabinol dimethylheptyl, 0.022 mg/kg], WIN 55,212-2 [(R)-4,5-dihydro-2-methyl-4(4-morpholinylmethyl)-1-(1-naphthalenylcarbonyl)-6H-pyrrolo [3,2,1-ij] quinolin-6-one, 0.41 mg/kg] and (-)-Delta9-tetrahydrocannabinol (0.41 mg/kg). Non-cannabinoids with analgesic properties, such as morphine, amitriptyline, carbamazepine, gabapentin and baclofen, did not generalize to the cue. It is concluded that the discriminative stimulus effects of BAY 59-3074 are specifically mediated by cannabinoid CB1 receptor activation [3].

Radioligand Displacement Assay [2]
Further characterization of 21 was performed using radioligand displacement of [3H]CP55940 and equilibrium dissociation constant (Ki) value was determined as described previously. Data reported are average values from 3 measurements with <30% standard error.

cAMP Accumulation Assay [2]
The cAMP assays were performed in Chinese Hamster Ovary (CHO) cells stably expressing the human CB1 receptor (hCB1) cultured under standard cell culture conditions (37 °C, 5% carbon dioxide, DMEM media with 1% Penicillin/Streptomycin and 400 µg/mL G418) using the Lance™ assay kit and manufacturer’s instructions were closely followed . In brief, stimulation buffer containing 1X Hank’s Balanced Salt Solution (HBSS), 5 mM HEPES, 0.1% BSA stabilizer, and 0.5 mM final IBMX was prepared and titrated to pH 7.4 at rt. Serial dilutions of the test compounds and 300 nM forskolin, both prepared at 4× the desired final concentration in stimulation buffer, were added to a 96-well white ½ area microplate. The CHO-hCB1 cells were lifted with a non-enzymatic solution, and 4000 cells were added to each well. After incubating for 30 min at room temperature, Eu-cAMP tracer and uLIGHT-anti-cAMP working solutions were added per the manufacturer’s instructions. After incubation for 1 h, the TR-FRET signal (ex 337 nm, em 620 and 650 nm) was read on a CLARIO star multimode plate reader. Data were analyzed using Prism software. Nonlinear regression analysis was performed to fit data and obtain maximum response (Emax), EC50, correlation coefficient (r2), and other parameters. All experiments were performed in duplicate 2–3 times to ensure reproducibility and data are reported as mean ± standard error of mean unless noted otherwise.

Male Wistar rats (160-250 g)
0.3 mg/kg, 1 mg/kg, and 3 mg/kg
Oral administration; daily; for 2 weeks.

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In rats, generalization of BAY 59-3074 to the cue induced by the cannabinoid CB(1) receptor agonist (-)-(R)-3-(2-hydroxymethylindanyl-4-oxy)phenyl-4,4,4-trifluoro-1-butanesulfonate (BAY 38-7271) in a drug discrimination procedure, as well as its hypothermic and analgesic effects in a hot plate assay, were blocked by the cannabinoid CB(1) receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride (SR 141716A). BAY 59-3074 (0.3-3 mg/kg, p.o.) induced antihyperalgesic and antiallodynic effects against thermal or mechanical stimuli in rat models of chronic neuropathic (chronic constriction injury, spared nerve injury, tibial nerve injury, and spinal nerve ligation models) and inflammatory pain (carrageenan and complete Freund's adjuvant models). Antiallodynic efficacy of BAY 59-3074 (1 mg/kg, p.o.) in the spared nerve injury model was maintained after 2 weeks of daily administration. However, tolerance developed rapidly (within 5 days) for cannabinoid-related side effects, which occur at doses above 1 mg/kg (e.g., hypothermia). Uptitration from 1 to 32 mg/kg p.o. (doubling of daily dose every 4th day) prevented the occurrence of such side effects, whereas antihyperalgesic and antiallodynic efficacy was maintained/increased. No withdrawal symptoms were seen after abrupt withdrawal following 14 daily applications of 1 to 10 mg/kg p.o. It is concluded that BAY 59-3074 may offer a valuable therapeutic approach to treat diverse chronic pain conditions.[1]

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Pharmacokinetic Testing [2]
Female C57BL/6 mice were bred in house and used at ~10 weeks of age for pharmacokinetic (PK) testing. Three animals were tested per time point. Doses were formulated in 2% NMP in canola oil, and all compounds were delivered at 3 mg/kg by intraperitoneal injection (IP). Tissues were taken at 0.5, 1, 2, and 4 h post dose. Animals were subjected to whole body perfusion using saline prior to tissue collection. Brain samples were homogenized with 50:50 ethanol:water (1:5, v/v). Forty µL of the homogenate, 10 µL of acetonitrile, and 150 µL of 100 ng/mL reserpine in acetonitrile containing 0.1% formic acid were vortexed and centrifuged. Plasma samples were diluted with 10 µL of acetonitrile, 150 µL of 100 ng/mL reserpine in acetonitrile containing 0.1% formic acid, vortexed and centrifuged. Samples were subjected to LC/MS/MS analysis. Standards were prepared in blank samples and used for calibration curves. Chromatography was performed using a Phenomenex Luna C18 column.

[1]. 3-[2-cyano-3-(trifluoromethyl)phenoxy]phenyl-4,4,4-trifluoro-1-butanesulfonate (BAY 59-3074): a novelcannabinoid Cb1/Cb2 receptor partial agonist with antihyperalgesic and antiallodynic effects. J Pharmacol Exp Ther. 2004 Aug;310(2):620-32.

[2]. Structure–Activity Relationship Development Efforts towards Peripherally Selective Analogs of the Cannabinoid Receptor Partial Agonist BAY 59-3074. Molecules. 2022 Sep 2;27(17):5672.

[3]. Discriminative stimulus effects of the structurally novel cannabinoid CB1/CB2 receptor partial agonist BAY 59-3074 in the rat. Eur J Pharmacol. 2004 Nov 28;505(1-3):127-33.

4,4,4-trifluoro-1-butanesulfonic acid is an aromatic ether.

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Selective modulation of peripheral cannabinoid receptors (CBRs) has potential therapeutic applications in medical conditions, including obesity, diabetes, liver diseases, GI disorders and pain. While there have been considerable efforts to produce selective antagonists or full agonists of CBRs, there has been limited reports on the development of partial agonists. Partial agonists targeting peripheral CBRs may have desirable pharmacological profiles while not producing centrally mediated dissociative effects. Bayer reported that BAY 59-3074 is a CNS penetrant partial agonist of both CB1 and CB2 receptors with efficacy in rat models of neuropathic and inflammatory pain. In this report, we demonstrate our efforts to synthesize analogs that would favor peripheral selectivity, while maintaining partial agonism of CB1. Our efforts led to the identification of a novel compound, which is a partial agonist of the human CB1 (hCB1) receptor with vastly diminished brain exposure compared to BAY 59-3074.[2]

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Structure–activity relationships of 1 (BAY 59-3074) would be useful in developing better compounds with limited CNS penetration. Recent crystal structures of hCB1 and docking studies with 1 indicate that the sulfonate linker is near to a polar region of the binding pocket and could be replaced with a polar hydrogen bond donating linker to facilitate development of peripherally selective analogs. Initial results showed this to be the case but controlling the level of efficacy is challenging. Effects on potency and efficacy are interdependent on the linker, the attached group, and the substitution pattern of the core. While a clear picture of the SAR to develop peripheral partial agonists of 1 did not emerge, we identified two hCB1 partial agonists with improved physical properties. Combining a change in the core substitution pattern from meta to para with replacement of the sulfonate linker with an amide resulted in the benzylamide 24 which has a hydrogen bond donor and is a partial agonist in the cAMP hCB1 assay. Replacement of the linker with a sulfamide and exchange of the alkyl chain with a cyclohexyl group resulted in the sulfamide 21, also a partial agonist in the cAMP hCB1 assay. Compound 21 already has physical properties that strongly favor peripheral selectivity (TPSA = 91, cLogP = 4.2, H donors = 2). This was confirmed in PK studies wherein brain concentration of this compound was greatly diminished versus that of 1. Additional SAR studies based on 21 and 24 may provide a map to compounds with the desired profile. The activity and efficacy of these compounds at hCB2 were not examined and will form the basis of future studies along with possible in vivo examination of effects.[2]

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3-[2-Cyano-3-(trifluoromethyl)phenoxy]phenyl-4,4,4-trifluoro-1-butanesulfonate (BAY 59-3074) is a novel, selective cannabinoid CB(1)/CB(2) receptor ligand (K(i) = 55.4, 48.3, and 45.5 nM at rat and human cannabinoid CB(1) and human CB(2) receptors, respectively), with partial agonist properties at these receptors in guanosine 5-[gamma(35)S]-thiophosphate triethyl-ammonium salt ([(35)S]GTPgammaS) binding assays. In rats, generalization of BAY 59-3074 to the cue induced by the cannabinoid CB(1) receptor agonist (-)-(R)-3-(2-hydroxymethylindanyl-4-oxy)phenyl-4,4,4-trifluoro-1-butanesulfonate (BAY 38-7271) in a drug discrimination procedure, as well as its hypothermic and analgesic effects in a hot plate assay, were blocked by the cannabinoid CB(1) receptor antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide hydrochloride (SR 141716A). [1]

C18H13NO4F6S

453.35552

453.047

C, 59.99; H, 5.03; F, 4.52; N, 26.65; O, 3.81

406205-74-1

10479060

White to off-white solid powder

1.496g/cm3

490.569ºC at 760 mmHg

250.488ºC

1.521

6.501

0

11

7

30

709

0

O=S(CCCC(F)(F)F)(OC1=CC=CC(OC2=CC=CC(C(F)(F)F)=C2C#N)=C1)=O

LWUSZIVDPJPVBW-UHFFFAOYSA-N

InChI=1S/C18H13F6NO4S/c19-17(20,21)8-3-9-30(26,27)29-13-5-1-4-12(10-13)28-16-7-2-6-15(14(16)11-25)18(22,23)24/h1-2,4-7,10H,3,8-9H2

[3-[2-cyano-3-(trifluoromethyl)phenoxy]phenyl] 4,4,4-trifluorobutane-1-sulfonate

BAY59-3074; BAY-593074; BAY-59-3074; BAY 59-3074; BAY 593074; BAY-59-3074; 1-Butanesulfonic acid, 4,4,4-trifluoro-, 3-(2-cyano-3-(trifluoromethyl)phenoxy)phenyl ester; BAY-593074; 5FO5Z101GU; CHEMBL1354658; 3-(2-Cyano-3-(trifluoromethyl)phenoxy)phenyl 4,4,4-trifluorobutane-1-sulfonate; BAY593074

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO: ~91 mg/mL (~200.7 mM)
Ethanol: ~91 mg/mL

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.51 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)