Chemoinformatic Analysis of Psychotropic And

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1 Chemoinformatic analysis of psychotropic and antihistaminic drugs in the light 2 of experimental anti-SARS-CoV-2 activities 3 4 Villoutreix BO1*, Krishnamoorthy R2, Tamouza R2, Leboyer M2, Beaune PH3, 5 6 7 8 1INSERM 1141, NeuroDiderot, Université de Paris, Hôpital Robert-Debré, F-75019 Paris, France

9 2Université Paris Est Créteil, INSERM U955, IMRB, Laboratoire Neuro-psychiatrie translationnelle, 10 AP-HP, Département Medico-Universitaire de Psychiatrie et d’Addictologie (DMU ADAPT), Hôpital 11 Henri Mondor, Fondation FondaMental, F-94010 Créteil, France

12 3INSERM U1138 Centre de recherche des Cordeliers 75006 Paris, and Université de Paris, Paris, France

14 * Corresponding author: [email protected]

27 28

30 Introduction: There is an urgent need to identify therapies that prevent SARS-CoV-2 infection 31 and improve the outcome of COVID-19 patients.

32 Objective: We proposed, before summer 2020, that cationic amphiphilic psychotropic and 33 antihistaminic drugs could protect psychiatric patients from SARS-CoV-2 infection based upon 34 clinical observations. At that time, experimental in vitro data on SARS-CoV-2 were missing.

35 Methods: Open high-throughput screening results are now available at the NCATS COVID19 36 portal and it is possible to investigate again our initial hypothesis using simple chemoinformatics 37 approaches but this time with in vitro data on SARS-CoV-2.

38 Results and Discussion: We here revisit our hypothesis in the light of SARS-CoV-2 experimental 39 screening results and propose that several cationic amphiphilic psychotropic and antihistaminic 40 drugs could possibly protect people from SARS-CoV-2 infection; some of these molecules have 41 very limited adverse effects and could eventually be used as prophylactic drugs. Further, recent 42 analysis of electronic health records reported by other research groups, including drug 43 combinations, also suggest that a small list of molecules could be of interest at different stages of 44 the disease progression. Taken together, these observations suggest that clinical trials are now 45 needed to fully evaluate the potentials of these molecules.

48 Keywords: chemoinformatics, Covid-19, high-throughput screening, phenothiazine, 49 prophylaxis

2 55 Introduction

56 Rapid pandemic spread of SARS-CoV-2 virus and the resulting COVID-19 has caused havoc

57 worldwide and continue to do so with transmission rate spikes. Hence there is an urgent need to

58 identify potential pharmacological prophylactic/therapeutic interventions both to prevent SARS-

59 CoV-2 infection and to improve the clinical outcome of COVID-19 patients. At present, only

60 vaccines offer hope but small molecules interfering with SARS-CoV-2 infection would have an

61 added value and could be cost effective. The shortest path to discover such compounds is via drug

62 repurposing approaches but it should be mentioned, as reported by Edwards1, that drug repurposing

63 seldom works in Human if there are no clinical observations as background rationale and/or in the

64 absence of a reasonable mechanistic hypothesis. Early 2020, we hypothesized, based upon clinical

65 observations made in several hospitals including the large psychiatric department of the Henri

66 Mondor hospital, Créteil (Paris area), France, and confirmed in several other hospitals, that

67 therapeutic molecules used in the treatment of psychiatric patients might have protected them from

68 SARS-CoV-2 infection and hence explained the paucity of admission to specialized COVID units.2

69 In order to gain knowledge about the putative molecules that could be repositioned against SARS-

70 CoV-2, we collected a list of 18 compounds most commonly prescribed in the in- and out-patient

71 Henri Mondor treatment programs (Alimemazine, Amisulpride, Aripiprazole, Cetirizine,

72 Citalopram, Clozapine, Cyamemazine, Diazepam, Escitalopram, Hydroxyzine, Lithium,

73 Lorazepam, Melatonin, Nicotine, Quetiapine, Sertraline, Valproate, and Zopiclone). Some of these

74 compounds belong to the phenothiazine class (e.g., Alimemazine and Cyamemazine), others

75 contain a benzodiazepine core (e.g., Lorazepam) or else are antihistamine drugs with, for example,

76 a core piperazine group (e.g., Cetirizine and Hydroxyzine). We investigated further these 18 drugs

77 using chemoinformatic strategies and via literature mining as it is well documented that some

3 78 molecules from these classes possess in vitro antiviral activities (reviewed in3). Indeed, ten out of

79 our 18 drugs have been reported to have in vitro antiviral activities against various types of viruses

80 including HIV, Ebola, HSV, MERS or SARS. Four were chemically close to molecules that

81 display such in vitro antiviral activity and were therefore predicted to also possess such property

82 based on “principle of similarity” commonly applied in medicinal chemistry. While the

83 psychotropic and antihistamine drugs that we were investigating belong to several chemical

84 families, we observed a common trend among these molecules. Eleven (60%) out of these 18 most

85 prescribed compounds were cationic amphiphilic drugs (CADs). CADs are molecules

86 characterized by the presence of a hydrophobic-aromatic ring system and a side chain that carries

87 one (or more) ionizable amine functional group(s) (i.e., a basic moiety). Many such compounds

88 accumulate in lysosomes or in other acidic subcellular compartments (i.e., lysosomotrophic agents,

89 a well-known molecule acting on SARS-CoV-2 in vitro at least via this mechanism, among others,

90 is the antimalarial drug hydroxychloroquine). Many CADs are therefore known to interfere with

91 intracellular trafficking (e.g., endosomal entry route, clathrin-mediated or not) and several such

92 molecules have indeed in vitro antiviral activity.4,5 CADs compounds are also known to induce

93 phospholipidosis in vitro (a lysosomal storage disorder), and 14 out of our list of 18 drugs were

94 known or predicted to induce phospholipidosis.6 These molecules are associated with intracellular

95 traffic perturbations and could play a role in SARS-CoV-2 infectivity.7

96 From these observations, presumably, many of our 18 molecules could play a role in the early

97 phases of virus life cycle. In short, early 2020, our hypothesis was that interesting compounds to

98 reposition against SARS-CoV-2 would be CAD psychotropic and antihistaminic molecules (i.e.,

99 the ones present in our list of 18 compounds or analogs that possess similar physicochemical

100 properties and chemical substructures, in our list of 18 compounds). These drugs could perturb

4 101 virus entry, for instance by interfering with normal endocytosis but could also inhibit directly or

102 indirectly the interaction between the viral Spike protein and the host ACE2 receptor (details about

103 cell entry mechanisms are reported here8). These drugs obviously bind to their primary targets (for

104 example histamine receptors, dopamine receptors, GABA(A) receptor) although the impact of

105 such interactions on COVID-19 is not clear. In addition, it is well-known that drugs can interact

106 with several targets and as such the CAD psychotropic and antihistaminic molecules are likely to

107 also bind to known (e.g., the Sigma receptors9) and unknown off-targets and act on the virus and/or

108 on the host defense mechanisms. The conclusion of our first analysis was that CAD psychotropic

109 and antihistaminic molecules could confer protection against SARS-CoV-2 infection in psychiatric

110 patients as these subjects receive as treatment a “CAD cocktail” that certainly interferes with the

111 endo-lysosomal viral machinery essential for cell entry, replication and spread to other cells2 while

112 some compounds (e.g., antihistamine drugs) could also down-regulate prominent inflammatory

113 messengers.4,10 Yet, in our first report, we also mentioned that while such drugs might protect

114 against SARS-CoV-2 infection, many of them do have concerns (e.g., toxic side effects if

115 unmonitored) and hence cannot be recommended for mass population prophylaxis and/or

116 treatment in the urgent fight against COVID-19.

117 When we reported our study about the possible protective potentials of psychotropic and

118 antihistamine drugs against SARS-CoV-2 infection2, in vitro data on SARS-CoV-2 were not

119 available. Since then, low-throughput and high-throughput screening experiments have been

120 performed (see a recent review11). To the best of our knowledge, the largest high-throughput

121 screening data open to the research/medical community is available at the NCATS COVID19

122 portal (https://opendata.ncats.nih.gov/covid19). These data involve the screening of over 10,000

123 chemical compounds. We here investigate our initial observations in the light of these new

5 124 experimental data. We thus further rationalize the potential protective effect of some psychotropic

125 and antihistamine drugs against SARS-CoV-2 infection and propose some already approved drugs

126 (with manageable/tolerable adverse effects) that could be repositioned to protect the general

127 population against COVID-19.

128

129 Methods

130 Data preparation and analysis

131 The first step involved the curation and preparation of the experimental screening data. We

132 downloaded the data on SARS-CoV-2 cytopathic effect (CPE) from the NCATS COVID19 portal.

133 This assay measures the ability of a compound to reverse the cytopathic effect induced by the virus

134 in Vero E6 host cells (one of the most commonly used cell lines for studying this family of virus).

135 The assay provides valuable basic information about the ability of a compound to restore the cells’

136 function but does not provide indication about the possible mechanism of action(s) of the small

137 chemical compound under study. We also explored using the same Vero E6 cells the cytotoxic

138 effect of these molecules in a counterscreen since the observed effect of some compounds may be

139 mediated through cell viability and could lead to erroneous interpretation of the CPE results. Such

140 information on cell viability is evidently important in the applicability of such compounds in

141 clinical pharmacological interventions. Some additional data are important to take into account in

142 the present analysis and involve the effect of the chemical compounds over human fibroblasts.

143 This assay measures the host cell ATP content as a readout for cytotoxicity. When available, we

144 annotated further the CPE data with the results of this assay. To be able to quickly repurpose a

145 compound, we focused essentially on molecules that have been approved or are in clinical trials

146 (in some instances drugs are only approved for veterinary use). We thus cross-linked the CPE

6 147 sample ID numbers to drug names (e.g., international nonproprietary names), PubChem SID or

148 CID numbers12 or to ChEMBL ID numbers13 and to molecules that possess ATC (Anatomical

149 Therapeutic Chemical Classification system) codes and/or have documented therapeutic

150 indications as found in DrugBank14, DrugCentral15 and eDrug3d16. Molecules that are at the

151 preclinical stage (e.g., chemical probes) were essentially excluded via this procedure. Additionally,

152 the salts, when present in the chemical file, were deleted using the Maya chemistry toolkit17 and

153 the chemistry of the molecules was standardized using our FAF-Drugs4 web server.18 The drugs

154 in duplicate were removed. Further curation of the CPE data also involved the following: i)

155 removal of a few molecules without potency values (AC50 values are reported for this dataset, i.e.,

156 concentration for half-maximal activity19), ii) investigation of the curve class (i.e., the evaluation

157 of the quality of the dose-response curve and thus the quality of the measurements, in the CPE

158 data, (a curve class of type 4 is a flat curve indicating no activity) and maximum response (i.e.,

159 maximum response value detected in the experiment). In such assay, compounds are usually

160 considered active when the maximum response (absolute) value is above 30.20 Further, we

161 removed molecules that were found highly cytotoxic in the Vero E6 counterscreen assay. From

162 the initial CPE data containing 10,721 assessed samples (some molecules have been screened

163 several times and most are chemical probes), we ended up with a list of 198 bioactive drugs. In

164 this list of drugs, 93 were not tested for toxicity on human fibroblast and for the remaining 105

165 molecules, only one was found cytotoxic (Disulfiram, used for alcohol addiction, this molecule

166 was also removed not only because of its cytotoxic nature in these assays but also as found to be

167 a promiscuous SARS-CoV-2 main protease inhibitor21). At this stage of the analysis, we decided

168 to retain all the compounds including those not annotated for human fibroblast toxicity. We then

169 focused our attention on families of molecules that contain several members and removed some

7 170 molecules with highly specific disease indication such as Deferasirox (an oral iron chelator given

171 to patients receiving long term blood transfusions). In this last step of data curation, we grouped

172 the 198 compounds according to chemistry and disease indications and removed molecules that

173 did not fulfill our selection criteria. This step led to the selection of a final list of 161 molecules.

174 Data were then visualized with DataWarrior.22 In this list, very few molecules are only chemical

175 probes, like Calpeptin. They were kept because they are often used to assess some enzymatic

176 activities. The structural classification of chemical entities was then performed using the

177 ClassyFire application.23

178

179 Results and Discussion

180 Our selected list of 161 bioactive drugs in the CPE assay (after excluding molecules that were

181 found to be highly cytotoxic in Vero E6 cells counterscreen assay) are prescribed as a treatment in

182 five major disease areas (Fig. 1). Of course, it is important to note that some of the selected drugs

183 can be given to different types of disease but we still attempted to group them into main/major

184 indications.

185 Molecules that are in the “Allergies” category (different types of allergies are involved) are here

186 all antihistamine compounds. Some of these molecules also have other properties such as anti-

187 inflammatory effects. This group of compounds include for instance Loratadine and Desloratadine

188 (e.g., allergic rhinitis), Azelastine (e.g., allergic and vasomotor rhinitis and allergic conjunctivitis)

189 and Promethazine (e.g., used for allergic reactions, nausea and vomiting). While not an

190 antihistamine molecule nor a CAD, Colchicine could be present in this group although it could

191 also be in the Cancers and Cardiovascular categories. It is an anti-inflammatory drug that could

192 reduce the risk of complications and death from COVID-19 (communication in January 2021 by

8 193 Dr. J.C. Tardif, Montreal Heart Institute). This molecule is not present in our list of 161 compounds

194 because it was not active in the CPE assays that we investigated and was cytotoxic. As such, this

195 molecule could possibly act on the exaggerated inflammatory response seen in some patients in

196 more advance stages of disease progression.

197

198 Another major group of drugs are anti-cancer agents. Many of these compounds are kinase

199 inhibitors such as Abemaciclib, Pazopanib, Ceritinib, Lapatinib and Imatinib. Imatinib, a potent

200 ABL inhibitor, was found here to have moderate CPE activity. This compound is in clinical trial

201 for COVID-19 but other authors noted that this molecule had limited activity on SARS-CoV-2

202 entry/infection in different in vitro assay types.24 It is also important to note that some kinase

203 inhibitors are used for the treatment of autoimmune disorders and other complex imbalance of the

204 immune system like for instance, Baricitinib. This compound is used for the treatment of

205 rheumatoid arthritis and is in clinical trial for treating COVID-19 patients. Some authors suggested

206 that it could be beneficial in preventing virus infectivity via perturbation of endocytosis25 and/or

207 could be important as inhibitor of targets in critically ill COVID-19 patients.26 This compound is

208 obviously not present in our list of anti-cancer agents and was not found active in the CPE assay.

209 Yet, such molecule could be of interest, for instance to act on the cytokine storm. Also, in the

210 cancer field, Pralatrexate was found able to potently inhibit SARS-CoV-2 replication with a

211 stronger inhibitory activity than Remdesivir.27 It is not present in our list as was found very

212 cytotoxic but in the in vitro assays that we analyzed. Yet, this compound could be of interest.

213

214 Several drugs prescribed in the field of cardiovascular diseases were also found active in the CPE

215 assay. These include Vorapaxar (reduces the incidence of thrombotic cardiovascular events or with

9 216 peripheral arterial disease, a molecule proposed as potentially interesting for the treatment of

217 COVID-19 patients28), Carvedilol (treatment of heart failure, hypertension, a molecule also

218 suggested in the literature as potentially beneficial for COVID-19 patients via several mechanisms

219 of action29) or Nicardipine (antihypertensive properties effective in the treatment of angina and

220 coronary spasms).

221

222 The molecules that belong to the “Infectious Disease” category include molecules with widely

223 different types of activities. For instance, Mefloquine and Chloroquine (anti-parasitics for

224 Malaria), Letermovir (for prophylaxis of cytomegalovirus infection), Cenicriviroc (under clinical

225 trials for the treatment of HIV-infection/AIDS), Nitazoxanide (broad-spectrum anti-infective drug)

226 and Lopinavir (in the treatment of HIV-infection). Several antimalarial drugs (e.g., Methylene

227 blue) are known to act in vitro on SARS-CoV-230 while the in vitro activity of Cenicriviroc has

228 also been recently reported.31 In the present study however, Methylene blue was not kept in our

229 final list of 161 compounds as the molecule was reported to be cytotoxic in the NCATS COVID19

230 results. Nitazoxanide was for example found promising in its ability to hinder the replication of a

231 SARS-CoV-2 isolate32 and is now in several clinical trials either alone (e.g., see33), against

232 placebo, or in combination with for instance Ivermectin (a broad-spectrum anti-parasite

233 medication that was not found to be active in the CPE assays that we investigated here and that

234 was highly cytotoxic in Vero E6 cells counterscreen).

235

236 Molecules of the “Psychotropic drugs” category also involve structurally diverse compounds that

237 include, for instance, Clomipramine (an antidepressant, also reported in a very recent study34),

10 238 Chlorpromazine (an antipsychotic that has activity in vitro and is in clinical trial35-37), Fluoxetine

239 (an antidepressant), and Haloperidol (an antipsychotic).

240

241 From these first analysis, it was then important to cluster the small molecules based on their

242 chemistry. Many different approaches can be used for this purpose38 but as the bioactive

243 compounds investigated here tend to belong to a limited number of chemical classes, a simple way

244 is to group the compounds based on the presence of identical/similar organic functional groups.

245 We investigated our list of 161 molecules using this strategy with the DataWarrior application.

246 The results are shown in Fig. 2. As can be seen, a large cluster involves drugs used for allergies

247 and for mental disorders. These molecules all share a phenothiazine group with different types of

248 side chains (molecules around Trimeprazine also named Alimemazine or around Promazine or

249 Fluphenazine, a piperazine ring phenothiazine). Other molecules that are chemically closely

250 related involve for instance Clomipramine or Imipramine. Here, the phenothiazine group is not

251 present but a dibenzazepine group is found instead.

252 Earlier we suggested that cationic amphiphilic psychotropic and antihistamine drugs might play a

253 significant protective role as witnessed by the relatively low representation of the psychiatric

254 patients among the COVID-19 clinical units.2 The notion of CADs and/or lysosomotropic active

255 agents is not clearly defined but in general, the scientific community considers that the cationic

256 character of a molecule can be flagged by the (predicted) value of its highest basic pKa while the

257 amphiphilicity can be estimated by (computed) log P values.4,39 Such compounds contain organic

258 bases with basic pKa above 6 or 7 and are expected to carry one or more positive charges at

259 physiological pH (even more so in the acidic environment of the endosomal-lysosomal system)

260 while the computed log P value of these molecules is in general above 3. If we apply such criteria

11 261 to our list of 161 molecules, 125 compounds have a computed log P above 3 and among these, 105

262 have at least one positive charge, suggesting that about 65% of the compounds could be of CADs.

263 Obviously such global physicochemical properties are present in a variety of drugs that act in many

264 therapeutic areas. Here we found such compounds in all the five disease categories reported in Fig.

265 1.

266

267 From our analysis, the types of chemical structures or substructures that would seem interesting to

268 interfere with SARS-CoV-2 infection are illustrated in Figure 3. According to the automated

269 structural classification package ClassyFire, the main chemical classes involve molecules that

270 contain as a core a phenothiazine group (e.g., Promazine), a benzazepine group (e.g., Imipramine),

271 a benzodiazepine group (e.g., Clozapine), a dibenzocycloheptene group (e.g., Cyproheptadine), a

272 benzocycloheptapyridine group (e.g., Desloratadine), a diazanaphthalene group (e.g., Azelastine)

273 or a benzene and substituted derivative group (e.g., Homochlorcyclizine or Buclizine, in this case

274 a piperazine ring is present in the core region).

275

276 In vitro data (and for this disease, small animal models) cannot clearly predict if a molecule is

277 going to be active on SARS-CoV-2 in Human. Yet, interestingly, after mining 219,000 digital

278 health records, Reznikov et al.40, have reported that prior usage of Loratadine, Diphenhydramine,

279 Cetirizine, Hydroxyzine or Azelastine was associated with a reduced incidence of positive SARS-

280 CoV-2 test results in individuals of 61 years and above. Loratadine and Azelastine are present in

281 our list of 161 drugs. Loratadine and Azelastine were also found in other reported experimental

282 screening data available at the open NCATS portal to perturb virus entry via modulation of the

283 ACE2-Spike protein interaction (similar results in this assay were apparently not found for

12 284 Desloratadine). Diphenhydramine was not tested in the data that we downloaded from the NCATS

285 portal but the highly similar molecule Bromodiphenhydramine was found active in the CPE assay.

286 Yet, Reznikov et al. reported that Diphenhydramine was indeed active in vitro. Hydroxyzine and

287 its active metabolite Cetirizine, were not found active in the CPE assays that we analyzed nor in

288 other assays reported at the open NCATS portal but Hydroxyzine was found to impede the

289 interaction between the viral Spike protein and the human receptor ACE2 and as such could

290 interfere with virus entry (action at the level of the Spike could also be of importance for SARS-

291 CoV-2 virulence). In our hands, Hydroxyzine has only moderate in vitro activity (unpublished

292 data) but we listed it as a very interesting CAD.2 In fact, very recent and interesting clinical

293 observations suggest that Hydroxyzine could reduce mortality in patients hospitalized for COVID-

294 19 most likely in part due to anti-inflammatory properties.41 These data suggest that additional

295 investigations are needed for this compound to fully understand the molecular mechanisms at play

296 and fully assess its activity on the virus and on the host. Along the same line and with the idea of

297 drug combination, it seems possible that the use of two anti-histamine molecules, Cetirizine and

298 Famotidine (not predicted to be CAD), both found to be inactive in the analyzed CPE assays, might

299 be a safe and effective approach to reduce the severity of COVID-19, presumably via a modulation

300 of the histamine-mediated cytokine storm.42 Again, one has to be cautious with in vitro data and

301 statistics performed on electronic health records, yet, the data reported above definitively suggest

302 that some of these CAD molecules could be of interest.

303 Overall, the present analysis, together with recently reported studies by other groups, support our

304 initial observation2 and do indeed suggest that “some” psychiatric patients could have been

305 protected by their pharmacological treatments. Along this line of reasoning, it should also be

306 mentioned that these patients receive in general a cocktail of drugs (psychotropic and anti-

13 307 histamine molecules) including several CAD compounds possibly working on different molecular

308 mechanisms.

309

310 Conclusions

311 From the above analysis, we suggest that some antihistamine and psychotropic CADs could protect

312 Humans from SARS-CoV-2 infection while some of these drugs might be also beneficial at more

313 advanced stages of the disease. Not only the recent publications mentioned above but also some

314 recent reviews suggest that more and more research teams in different countries believe, based on

315 different types of data, that these molecules could have a potential (e.g., some new reviews43-46).

316 The CAD molecules analyzed here have moderate to relatively high levels of anti-viral activity at

317 nontoxic concentrations in vitro. Some of these compounds are actively investigated such as

318 Fluoxetine (e.g., treatment of depression)47 or Chlorpromazine35,48 but many others need to be

319 explored. The molecular mechanisms involved are likely to be complex, from perturbation of

320 endocytosis, membrane fusion to direct binding to some specific receptors. For example, a recent

321 investigation applied surface plasmon resonance to study some antipsychotic drugs and reported

322 that several compounds could bind directly to ACE2 and partially inhibit this virus entry

323 mechanism.49 It is of course important to note that psychotropic compounds can induce side effects

324 and might not be safe enough to be prescribed for prophylaxis to a population not concerned by

325 psychiatric issues. Yet, with regard to SARS-CoV-2 infection, psychotropic CADs could be

326 replaced by antihistamine CADs. Clinical trials are now needed to investigate the true potentials

327 of these molecules in Human, the type of patients that could benefit from these molecules and at

328 which stage of the infection the drug(s) should be given, prophylaxis being a potential option. In

329 fact, for prevention purposes, molecules that have moderate in vitro activity might be sufficient

14 330 while it is also known, that, if needed, most antihistamine CADs could be tolerated at higher doses

331 without inducing severe side effects. Most likely, drug combinations could be beneficial but here

332 also clinical trials will be needed to evaluate potential additivity, synergy or antagonism. Further,

333 these compounds, if active in Human, may help facing the emergence of potentially devastating

334 variants while new vaccines are developed.

335 336 337 338 Contributors 339 BV planed, generated, analyzed the chemical data and wrote a first draft of the manuscript. ML 340 provided clinical observations. BV, RK and PB optimized the initial draft; a final version was 341 produced by all authors. 342 343 344 Declaration of Competing Interest 345 The authors declare that they have no competing financial interests or personal relationships that 346 could influence the work reported in this paper.

347 Acknowledgements 348 We thank the foundation FondaMental for supports.

349 350 351 352 Figure legend 353 Fig. 1: Five major drug indications found to have SARS-CoV-2 cytopathic effect 354 355 356 Fig. 2: Bioactive compounds (161 molecules) clustered according to the presence of organic 357 functional groups. The molecules that were found to share identical or similar organic functions 358 are connected by lines (eg., the presence of a specific ring system). Increased transparency of the 359 lines reflects decreasing similarity. The compounds are marked either as a circle (molecules not 360 toxic in human fibroblast toxicity assay) or as a square (molecules not tested for such toxicity). 361 The marker symbols and the connecting lines are color coded according to the reported 362 experimental potency values (for this dataset, AC50 values are provided). The background 363 colors behind the markers and lines are color coded according to the disease categories. Yet, as 364 some compounds in the category of “Allergies” are also prescribed in psychiatric settings 365 (indeed some psychotropic drugs were initially used as antihistaminics), we applied the same 366 background color for these two categories to facilitate the reading of this figure. We can observe 367 in this figure a large group of compounds sharing similar substructures (e.g., around

15 368 Promethazine) that have in general moderate activity in these in vitro assays.

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