Compd ARYL n % act. at 1E-06 M

SERT D2 5-HT7 5-HT1A M3 hERG

1 0 100 97 94 97 1

2 0 98 97 95 99 10 6

3 0 98 97 94 97 0 12

4 0 100 99 96 100 10 11

5 0 98 98 97 98 8 8

6 0 99 99 99 100 8 12

7 0 99 100 100 99 3 9

8 0 100 100 102 99 10 0

9 2 99 99 88 99 4

10 2 101 99 87 100 9

11 2 101 100 90 98 9

12 2 101 100 90 97 0 3

13 2 101 101 93 100 11

15 2 101 100 89 99 11

16 2 101 100 92 99 10 19

Marcin Kołaczkowski1, Monika Marcinkowska 1, Adam Bucki 1, Joanna Śniecikowska 1, Maciej Pawłowski 1, Grzegorz Kazek 1,Agata Siwek 1, Anna Wesołowska 1, Magdalena Jastrzębska-Więsek 1, Anna Partyka 1,

Paweł Mierzejewski 3, Przemyslaw Bienkowski 3

Novel ligands acting as SERT blockers and dopamine D2 receptor partial agonists with therapeutic potential for treatment of mood disorders and their comorbidities

1Faculty of Pharmacy, Jagiellonian University Medical College, Cracow, Poland; 2Adamed Ltd., Pieńków, Poland 3Institute of Psychiatry and Neurology, Warsaw, Poland

marcin.kolaczkowski@uj.edu.pl

Studies were financed by Adamed Ltd, Pieńków, Poland and National Center for Research and Development (NCBiR), Warsaw, Poland

Target pKi

SERT 8.2

D2 8.4

5-HT7 7.0

5-HT1A 7.7

M3 <6

5-HT2C <6

hERG <6

Mood disorders

Chemical structure of the lead molecule ADN-3662

Saccharine self-administration MED = 3 mg/kg

Reversal of haloperidol-induced deficits in saccharine self-administration active dose = 1 mg/kg

Mood (affective) disorders are a wide and heterogeneous group of medical conditions including unipolar and bipolar depression, mania or substance-induced mood disorders.

Their typical comorbidities include eating disorders, anxiety disorders or drug addiction.

Anhedonia and affective flattening are also major features of negative symptoms of schizophrenia [1]

Selective serotonin reuptake inhibitors (SSRI) acting through blockade of serotonin transporter (SERT) are a frequently-employed therapeutic options in various affective disorders [2].

On the other hand, dopamine D2 receptor partial agonists (e.g. aripiprazole), developed primarily as novel antipsychotic drugs, have attracted increasing interest also as promising mood modulating agents [3] and proved synergy with SSRIs [4,5].

Nevertheless, the therapeutic efficacy of current drugs remains limited and mood disorders present numerous unmet needs.

Series of novel arylsulfonamides

We designed and synthesized a novel series of arylsulfonamide derivatives, preferentially targeting 5-HT7 receptors

The group containing over 300 compounds was protected by an international patent application WO2012035123 [6]

General formula of the subset simultaneously targeting 5-HT7 as well as SERT, D2, and 5-HT1A receptors is presented below:

D2/5-HT1A

SERT 5-HT7 receptor

In vitro profile of ADN-3662

Significantly active in well validated mood deficit model

Selected in vivo data for ADN-3662

Active in a model of secondary negative symptoms

Active in other well-recognized models of mood deficits

Additional anxiolytic effects

Very wide therapeutic window – unwanted activities (catalepsy, sedation, memory imparement) not observed up to doses 10-100x higher than the effective ones.

No activities associated with side-effects: anticholinergic properties, hERG blockade etc.

Pharmacological profile of ADN-3662 warrants its further development in the treatment of mood deficits and related

comorbidities

Series of arylsufonamide derivatives displayed strong SERT, D2, 5-HT7 and 5-HT1A receptor affinity with reduced activity at undesirable molecular targets, notably M3 receptors and hERG channels

Based on in vitro and in vivo data compound 12 was chosen as the lead molecule for further evaluation. It’s code name is ADN-3662

Blockade of SERT and 5-HT7R as well as partial agonism at D2R and 5-HT1AR

provide synergistic effects in mood improvement

Series of arylsulfonamide derivatives was designed to specifically interact with SERT and 5-HT7 receptors

Crucial role of arylsulfonamide moiety for interaction with 5-HT7 binding sites

Sulfonamide oxygen atoms form specific H-bonds with Arg7.36 side chain

Aryl moiety penetrates hydrophobic pocket in region of transmembrane helices (TMH) 1, 2 and 7

Tetrahydropiridineindole (THPI) moiety provides affinity for serotonin transporter (SERT) anchoring at Asp98 and having hydrophobic contacts with Tyr176, Phe95 and Val343

Arylsulfonamide group additionally stabilises the complex by H-bond with Trp103 and cation-pi contact with Arg104

Structures and in vitro data for the most interesting examples are collected in Table 1:

[1] Der-Avakian A, Markou A. Trends Neurosci. 2012 Jan;35(1):68-77

[2] Blier P, El Mansari M., Philos Trans R Soc Lond B Biol Sci. 2013 Feb 25;368(1615): 20120536.

[3] Brown R, Taylor MJ, Geddes J. Cochrane Database Syst Rev. 2013 Dec 17;12:CD005000

[4] Worthington JJ, 3rd, Kinrys G, Wygant LE, Pollack MH. Int Clin Psychopharmacol 2005; 20(1): 9-11..

[5] Sheffrin M, Driscoll HC, Lenze EJ, et al. . J Clin Psychiatry 2009; 70(2): 208-13

[6] Kolaczkowski, M.; Kowalski, P.; Jaskowska, J.; Marcinkowska, M.; Bucki, A.; Wesolowska, A.; Pawlowski, M.; ‘Arylsylfonamides for the treatment of CNS diseases’ – international patent application, priority date: 2010.09.17, international filing date: 2011.09.16, publication date: 2012.03.22. WO 2012/035123

References

pKi

SERT

5-HT7 D2

5-HT1A hERG

M3 5-HT2C

5-HT1A D2 receptor

Molecular modeling showed also complementarity with D2 and 5-HT1A receptors

5-HT7 SERT

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0.0 mg/kg 1.0 mg/kg 3.0 mg/kg 10.0 mg/kg

ADN 3662 dose

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ADN 3662 dose

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Test procedure MED mg/kg Porsolt’s forced swim test in mice 1.25 Tail suspension test in mice 0.312 Porsolt’s forced swim test in rats 1 Four-plate test in mice 0.312 Expression of saccharine self-administration in rats 3 Vogel’s conflict drinking test 1 Reversal of haloperidol-induced deficits in saccharine self-administration in rats 1

Catalepsy in rats >100 Sedation in rats >100 Disruption of social behavior in rats >30

Sulfonamide oxygen atoms interact with Asn7.39 while aryl moiety interacts with aromatic cluster formed by Tyr2.64 and Trp7.40 in 5-HT1A binding sites

The NH group of THPI moiety stabilises the complex with D2 receptor by forming additional H-bond with Ser5.42