Wai N. Chan

1,3-Biarylureas as Selective Non-peptide Antagonists of the Orexin-1 Receptor

This communication reports SARs for the first orexin-1 receptor antagonist series of 1-aryl-3-quinolin-4-yl and 1-aryl-3-naphthyridin-4-yl ureas. One of these compounds, 31 (SB-334867), has excellent selectivity for the orexin-1 receptor, blood-brain barrier permeability and shows in vivo activity following ip dosing.

Identification of (-)-cis-6-acetyl-4S-(3-chloro-4-fluoro-benzoylamino)- 3,4-dihydro-2,2-dimethyl-2H-benzo[b]pyran-3S-ol as a potential antimigraine agent.

Optimisation of novel cis- and trans-4-(substituted-amido)benzopyran-3-ol derivatives has led to the identification of SB-220453 20 with an in vivo pre-clinical CNS profile predictive of potential antimigraine activity.

Profile of SB-204269, a mechanistically novel anticonvulsant drug, in rat models of focal and generalized epileptic seizures

1 Earlier optimization of structure‐activity relationships in a novel series of 4‐(benzoylamino)‐benzopyrans, led to the discovery of SB‐204269 (trans‐(+)‐6‐acetyl‐4S‐(4‐fluorobenzoylamino)‐3,4‐dihydro‐2,2‐dimethyl‐2H‐benzo[b]pyran‐3R‐ol, hemihydrate), a potent orally‐active anticonvulsant in the mouse maximal electroshock seizure threshold (MEST) test. 2 Studies have now been undertaken to determine the effects of SB‐204269 in a range of seizure models and tests of neurological deficits in rats. In addition, the compound has been evaluated in a series of in vitro mechanistic assays. 3 SB‐204269 proved to be an orally‐effective anticonvulsant agent, at doses (0.1–30 mg kg−1) devoid of overt behavioural depressant properties, in models of both electrically (MEST and maximal electroshock (MES)) and chemically (i.v. pentylenetetrazol (PTZ) infusion)‐evoked tonic extension seizures. However, the compound did not inhibit PTZ‐induced myoclonic seizures at doses up to 30 mg kg−1, p.o. 4 SB‐204269 also selectively reduced focal electrographic seizure activity in an in vitro elevated K+ rat hippocampal slice model at concentrations (0.1–10 μM) that had no effect on normal synaptic activity and neuronal excitability. 5 In all of these seizure models, SB‐204269 was equivalent or better than the clinically established antiepileptic drugs carbamazepine and lamotrigine, in terms of anticonvulsant potency and efficacy. 6 Unlike SB‐204269, the corresponding trans 3S,4R enantiomer, SB‐204268, did not produce marked anticonvulsant effects, an observation in accord with previous findings for other related pairs of trans enantiomers in the benzopyran series. 7 In the rat accelerating rotarod test, a sensitive paradigm for the detection of neurological deficits such as sedation and motor incoordination, SB‐204269 was inactive even at doses as high as 200 mg kg−1, p.o. This was reflected in the excellent therapeutic index (minimum significantly effective dose in the rotarod test/ED50 in the MES test) for SB‐204269 of >31, as compared to equivalent values of only 7 and 13 for carbamazepine and lamotrigine, respectively. 8 At concentrations (10 μM) well above those required to produce anticonvulsant activity in vivo (i.e. 0.1 μM in brain), SB‐204269 did not interact with many of the well known mechanistic targets for established antiepileptic drugs (e.g. Na+ channels or GABAergic neurotransmission). Subsequent studies have shown that the anticonvulsant properties of SB‐204269 are likely to be mediated by a novel stereospecific binding site present in the CNS. 9 The overall efficacy profile in rodent seizure models, together with a minimal liability for inducing neurological impairment and an apparently unique mechanism of action, highlight the therapeutic potential of SB‐204269 for the treatment of refractory partial and generalized tonic‐clonic seizures.

Characterization of the binding of [3H]‐SB‐204269, a radiolabelled form of the new anticonvulsant SB‐204269, to a novel binding site in rat brain membranes

1 SB‐204269 (trans‐(+)‐6‐acetyl‐4S‐(4‐fluorobenzoylamino)‐3,4‐dihydro‐2,2‐dimethyl‐2H‐benzol[b]pyran‐3R‐ol, hemihydrate) shows potent anticonvulsant activity in a range of animal seizure models, with a lack of neurological or cardiovascular side‐effects. The profile of the compound suggests that it may have a novel mechanism of action. This study describes the characteristics of a binding site for [3H]‐SB‐204269 in rat forebrain membranes. 2 Specific [3H]‐SB‐204269 binding was saturable and analysis indicated binding to a homogenoeous population of non‐interacting binding sites with a dissociation constant (KD) of 32±1 nM and a maximum binding capacity (Bmax) of 253±18 fmol mg−1 protein. Kinetic studies indicated monophasic association and dissociation. Binding was similar in HEPES or Tris‐HCl buffers and was unaffected by Na+, K+, Ca2+ or Mg2+ ions. Specific binding was widely distributed in brain, but was minimal in a range of peripheral tissues. 3 Specific [3H]‐SB‐204269 binding was highly stereoselective, with a 1000 fold difference between the affinities of SB‐204269 and its enantiomer SB‐204268 for the binding site. The affinities of analogues of SB‐204269 for binding can be related to their activities in the mouse maximal electroshock seizure threshold (MEST) test of anticonvulsant action. 4 None of the standard anticonvulsant drugs, phenobarbitone, phenytoin, sodium valproate, carbamazepine, diazepam and ethosuximide, or the newer anticonvulsants, lamotrigine, vigabatrin, gabapentin and levetiracetam, showed any affinity for the [3H]‐SB‐204269 binding site. A wide range of drugs active at amino acid receptors, Na+ or K+ channels or various other receptors did not demonstrate any affinity for the binding site. 5 These studies indicate that SB‐204269 possesses a specific CNS binding site which may mediate its anticonvulsant activity. This binding site does not appear to be directly related to the sites of action of other known anticonvulsant agents, but may have an important role in regulating neuronal excitability.

Stereochemical differentiation of anticonvulsant and antihypertensive effects in 4-(fluorobenzoylamino)-benzopyrans

Abstract Replacement of the 2-pyrrolidinone group of cromakalim 1 by the fluorobenzoylamino group has introduced anticonvulsant activity. It is particularly noteworthy that this activity is found predominantly in the 3R,4S enantiomeric series. In contrast, antihypertensive activity is restricted to the 3S,4R enantiomeric series.

Identification of a series of 1,2,3,4-tetrahydroisoquinolinyl- benzamides with potential anticonvulsant activity

A series of N-(tetrahydroisoquinolinyl)-2-methoxybenzamides was identified by high-throughput screening at the novel SB-204269 binding site. SAR studies have provided compounds 4 and 14 with high affinity and good anticonvulsant activity in animal models.

Evaluation of a series of anticonvulsant 1,2,3,4-tetrahydroisoquinolinyl-benzamides.

SAR studies around a series of N-(tetrahydroisoquinolinyl)-2-methoxybenzamides, identified by high-throughput screening at the novel SB-204269 binding site, have provided compounds such as 13, 29-33 with high affinity and excellent anticonvulsant activity in animal models.

Conformational preference of the 6-acetyl group in novel anticonvulsant trans 4S-benzamido-benzo[b]pyran-3R-ols

Abstract Conformationally restricted ketones derived from the novel anticonvulsant 4 S (4-fluorobenzoylamino)benzopyran SB-204269 1 have revealed a preferred “in plane” conformation which is essential for optimal potency at a unique receptor site.

Stereochemical preferences and requirement for the 3-hydroxyl group in novel anticonvulsant 4-fluorobenzoylamino benzopyrans

Abstract A cis 3 S ,4 S isomer, derived stereospecifically from an anticonvulsant trans 3 R ,4 S -( para -fluorobenzoylamino)-benzopyran using the DAST reagent, has been shown to possess anticonvulsant properties. In contrast the cis 3 R ,4 R enantiomer did not possess anticonvulsant properties but caused blood pressure to fall.

Use of the radioligand [125I]-SB-217644 in the characterisation and solubilisation of the novel binding site for the anticonvulsant SB-204269 in rat brain membranes and cell lines

Abstract SB-204269 (trans-(+)-6-acetyl-4S-(4-fluorobenzoylamino)-3,4-dihydro-2,2-dimethyl-2H-benzo[b]pyran-3R-ol) shows anticonvulsant activity in a range of animal seizure models, with a high therapeutic index and a lack of side-effects. We have previously reported the characterisation of a novel binding site for [3H]-SB-204269 in rat forebrain, which has a unique profile unrelated to other known anticonvulsant sites of action. We now describe the use of a [125I]-labelled form of SB-217644 (trans-6-acetyl-4S-(3-iodobenzoylamino)-3,4-dihydro-2,2-dimethyl-2H-benzo[b]pyran-3R-ol), an analogue of SB-204269, for studies on this novel binding site. In rat forebrain membranes, [125I]-SB-217644 shows a similar binding profile to that of [3H]-SB-204269, with a maximum specific binding capacity (Bmax) of 286±12 fmol/mg protein, but has twenty-fold higher affinity (Kd value 1.7±0.1 nM). The high affinity and high specific activity of [125I]-SB-217644 allowed it to be used for detection and characterisation of the detergent-solubilised form of the binding site. Specific [125I]-SB-217644 binding to cholate-solubilised rat cerebellum showed a Kd value of 2.7±0.3 nM and a Bmax value of 55±11 fmol/mg protein, with a 7.3±0.3% yield of solubilised binding sites. [125I]-SB-217644 was also used in whole-cell binding assays for investigation of the properties of the novel binding site in a range of cell lines. Both rat brain neuronal and glial primary cultures and several CNS-related cell lines were found to have levels of specific [125I]-SB-217644 binding similar to those present in rat forebrain membranes. The solubilisation of this novel binding site, and the ability to quantify and characterise it in solubilised tissues and whole cells using [125I]-SB217644, will allow further studies towards the ultimate identification of the molecular target of SB-204269.