Available online at www.jbr-pub.org.cn

Open Access at PubMed Central

The Journal of Biomedical Research, 2019 0(0): 1–6

Supplementary Data

Identification of therapeutic drugs against COVID-19 through computational investigation on drug repurposing and structural modification

Yangfang Yun1, Hengyi Song1, Yin Ji2, Da Huo1, Feng Han1, Fei Li1, Nan Jiang1, ✉

1Key Laboratory of Cardiovascular & Cerebrovascular Medicine, School of Pharmacy, Nanjing Medical University, Nanjing, Jiangsu 211166, China; 2The State Key Laboratory of Translational and Innovative Drug Development, Simcere Pharmaceutical Group, Nanjing, Jiangsu 210042, China.

O O O NH O O O P O O O 2 O N HN O N O P OO N O O N N N HO OH N N

H2N Tenofovir (9.2075) Remdesivir (8.9948) N

N NH2 H2N NH2 O N O O O N N N OH H O OH H O HN N O NH O N O NH2 N N N N N O N HO N N O NH2 H N O H O H

Atazanavir (8.9444) Va acyc ovir (8.2404) Va gancic ovir (7.4403) Abacavir (7.2371)

O O O F O F O O P O O N F S HN O OH O O OH N N N O H H H N N NH N N N O S O O N N N N N O H O O H O C N N N H2N Doravirine (7.2198) Adefovir (7.1527) Lopinavir (7.0424) Ritonavir (7.0171)

Supplementary Fig. 1 Structural formulas of the screened antiviral compounds and their docking scores at spike-ACE2 interface.

✉Corresponding author: Nan Jiang, Key Laboratory of Cardio- CLC number: R966, Document code: A vascular &Cerebrovascular Medicine, School of Pharmacy, The authors reported no conflict of interests. Nanjing Medical University, 101 Longmian Avenue, Nanjing, Jiangsu 211166, China. Tel/Fax: +86-25-86868485/+86-25- This is an open access article under the Creative Commons Attribu- 86868467, E-mail: [email protected]. tion (CC BY 4.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the Received 01 April 2020, Revised 22 June 2020, Accepted 30 June original work is properly cited. 2020, Epub 31 August 2020

© 2019 by the Journal of Biomedical Research. https://doi.org/10.7555/JBR.34.20200044 2 Yun Y et al. J Biomed Res, 2019, 0(0)

AB ACE2 ACE2

SubunitⅠ

SubunitⅡ SubunitⅢ SubunitⅢ

Spike Spike

Supplementary Fig. 2 Three-dimensional structure of spike-ACE2 complex. A: Spike protein contains three subunits, Ⅰ, Ⅱ, and Ⅲ, which are colored in blue, green, and purple, respectively. B: Only subunit Ⅲ of spike is exhibited and the active pocket locating at the inter- face between spike and ACE2 is shown in gray.

40 ETOT EPTOT EKTOT 20

0

−20

−40 Remdesivir 20 N6 H29 H30 kcal/mol) −3 0 OE OG

−20

−40 Energy (×10 Energy Glu17@ACE2 Ser480@spike 20

0 Supplementary Fig. 5 The atomic index of remdesivir and its interacting residues. −20

−40 0 5 10 15 20

Simulation time (nanosecond) Tenofovir

Supplementary Fig. 3 Fluctuations of total (ETOT), potential energies (EPTOT), and kinetic (EKTOT) of the studied protein- ligand complexes with the simulation time. O9 H N H29

N3 Remdesivir Tenofovir Lopinavir OE1 0 Phe476@spike Gln479@spike −10

−20

−30

(kcal/mol) Tenofovir

Binding energy N2 H −40 H29 H30 11−15 nanosecond N N3 11−20 nanosecond −50 O ND1 Ser480@spike Hie16@ACE2 Supplementary Fig. 4 Binding energies of studied compounds to spike-ACE2 coupling complex calculated from different simu- Supplementary Fig. 6 The atomic index of tenofovir and its in- lation time. teracting residues. Computational identification of candidate anti-SARS-CoV-2 drugs 3

A 16 Lopinavir 12 (Å) 4 Tyr491@spike 8 Ave: 4.21 Å O1 d4 4

H N4 Distance, d N N33 O4 0 0 2 4 6 8 10 Tenofovir Ser480@spike HZ OE Simulation time (nanosecond) N Glu17@ACE2 B 16

(Å) 12

5 Tenofovir Ave: 4.44 Å Lys13@ACE2 8 d5 4

Supplementary Fig. 7 The atomic index of lopinavir and its Distance, d interacting residues. 0 0 2 4 6 8 10 Hie16@ACE2 Simulation time (nanosecond)

Supplementary Fig. 9 Hydrophobic interaction between teno- fovir and spike-ACE2.

A 16 A 16 Lopinavir

(Å) 12 Remdesivir 1

(Å) 12 Ave: 4.72 Å 6 Ave: 4.45 Å 8 d1 8 4

Distance, d 4 d6 0 Distance, d 0 2 4 6 8 10 Phe476@spike 0 0 2 4 6 8 10 Simulation time (nanosecond) Tyr435@spike Simulation time (nanosecond) B 16 B 16 Remdesivir Lopinavir

(Å) 12

(Å) 12 2 7 Ave: 4.81 Å Ave: 4.67 Å 8 8 d 4 d1 4 7 Distance, d Distance, d 0 0 0 2 4 6 8 10 Lys13@ACE2 0 2 4 6 8 10 Phe476@spike Simulation time (nanosecond) Simulation time (nanosecond) C 16 C 16 Lopinavir Remdesivir (Å) 12 (Å) 12 3 8 Ave: 5.22 Å Ave: 4.58 Å d 8 3 8

4 4 Distance, d Distance, d d Phe54@ACE2 8 0 0 0 2 4 6 8 10 0 2 4 6 8 10 Hie16@ACE2 Simulation time (nanosecond) Simulation time (nanosecond)

Supplementary Fig. 8 Hydrophobic interaction between rem- Supplementary Fig. 10 Hydrophobic interaction between lop- desivir and spike-ACE2. inavir and spike-ACE2. 4 Yun Y et al. J Biomed Res, 2019, 0(0)

Supplementary Table 1 Potential drugs obtained through high-throughput screening Name Score Name Score

Tenofovira 9.2075 Abacavira 7.2371

Remdesivirb 8.9948 Doravirinea 7.2198

Atazanavira 8.9444 Adefovira 7.1527

Valacyclovira 8.2404 Lopinavira 7.0424

Valganciclovira 7.4403 Ritonavira 7.0171 Deferoxamine mesylate 12.8960 Naloxegol oxalate 8.7217 Goserelin acetate 10.6346 Octenidine dihydrochloride 8.6713 Landiolol hydrochloride 10.5632 Ethaverine hydrochloride 8.6713 Roxatidine acetate hydrochloride 10.2444 Carvedilol 8.6469 Bimatoprost 10.0981 Mitoxantrone hydrochloride 8.6327 Orlistat 10.0383 n-docosanol 8.5666 Naringin 10.0295 Tirofiban hydrochloride monohydrate 8.5228 Verapamil hydrochloride 10.0185 Esmolol hydrochloride 8.5129 Daurisoline 9.8525 Ethambutol dihydrochloride 8.4801 Tazemetostat 9.8259 Risedronic acid 8.4645 Leuprolide acetate 9.7606 Tianeptine sodium 8.4628 Coenzyme Q10 9.7268 Risedronate sodium 8.4608 Aspartame 9.7041 Ixazomib 8.4296 Thymopentin 9.4585 Glutathione 8.4278 Miriplatin 9.3745 Miltefosine 8.4256 Atorvastatin calcium 9.3730 Enzastaurin 8.4103 Flunixin meglumine 9.3568 DL-α-tocopherol 8.3863 Dabigatran etexilate 9.2073 Cediranib 8.3650 Batilol 9.1369 Mycophenolic acid 8.3294 Salvianolic acid B 9.0966 Methotrexate 8.3198 Batyl alcohol 9.0465 D-sorbitol 8.3188 Carfilzomib 8.9885 Visomitin 8.3188 Fulvestrant 8.8875 D-Mannitol 8.3188 Azelnidipine 8.8552 Pentagastrin 8.3182 Dronedarone hydrochloride 8.8402 Erythromycin 8.3108 Mitoxantrone 8.8016 Menatetrenone 8.2955 Prostaglandin E1 8.7995 Isavuconazonium sulfate 8.2915 Fingolimod hydrochloride 8.7727 Acrivastine 8.2890 Vitamin K2 8.2581 Lapatinib ditosylate monohydrate 7.7965 Cetylpyridinium chloride monohydrate 8.2440 Otilonium bromide 7.7900 Propafenone 8.2364 Dofetilide 7.7876 Amlodipine 8.2266 gamma-Linolenic acid 7.7805 Vitamin E 8.2161 Lercanidipine hydrochloride 7.7800 Ibutilide Fumarate 8.2124 Oxytocin acetate 7.7756 Lercanidipine 8.1933 Manidipine dihydrochloride 7.7664 Mupirocin 8.1786 Vardenafil hydrochloride trihydrate 7.7560 Montelukast sodium 8.1648 Suplatast tosilate 7.7522

Osimertinib mesylate 8.1647 mbelin 7.7438 Computational identification of candidate anti-SARS-CoV-2 drugs 5

Supplementary Table 1 Potential drugs obtained through high-throughput screening (continued) Name Score Name Score Sivelestat 8.1632 Lisinopril dehydrate 7.7287 Dyclonine hydrochloride 8.1300 (±)-Bisoprolol hemifumarate 7.7273 palovarotene 8.1290 Docusate sodium 7.7250 Fluprostenol isopropyl ester 8.0621 Prostaglandin E2 7.7008 Citicoline 8.0589 LCZ696 7.6718 Nadolol 8.0536 Indacaterol 7.6697 Zolmitriptan 8.0204 L-Arginine 7.6623 Diminazene Aceturate 8.0058 Pramoxine hydrochloride 7.6531 Amikacin 7.9969 Heptaminol hydrochloride 7.6518 Tamsulosin hydrochloride 7.9640 Mebeverine hydrochloride 7.6452 Dabigatran 7.9172 L-(-)-α-Methyldopa 7.6406 Plerixafor 8HCl 7.9115 Chidamide 7.6273 Lapatinib 7.8904 Flunixin meglumin 7.6261 Cetrimonium bromide 7.8824 Tandutinib 7.6155 Dibucaine 7.8780 D-Pantethine 7.6145 Pretomanid 7.8576 Erlotinib 7.6059 Dapagliflozin 7.8537 Cetilistat 7.6035 Nifekalant hydrochloride 7.8447 Deferasirox 7.5987 Etilefrine hydrochloride 7.5891 Nimodipine 7.4707 Iloperidone 7.5832 Zinc undecylenate 7.4347 Efonidipine 7.5794 Cilastatin 7.4163 Bambuterol hydrochloride 7.5761 MLN9708 7.4107 Benserazide hydrochloride 7.5721 Benactyzine hydrochloride 7.3977 Sumatriptan succinate 7.5497 Gastrodin 7.3975 Betaxolol hydrochloride 7.5422 Euquinine 7.3956 Menadione 7.5321 Spiramycin 7.3938 Procainamide hydrochloride 7.5212 Quinapril hydrochloride 7.3825 Fluvastatin sodium salt 7.5208 Proadifen hydrochloride 7.3821 Lumefantrine 7.5192 Manidipine 7.3672 Alprenolol 7.5171 Nefazodone hydrochloride 7.3635 Glafenine 7.5130 Flavin mononucleotide 7.3628 Sulfadimethoxine 7.5038 Pemetrexed disodium 7.3591 Sodium cromoglycate 7.4999 Folic acid 7.3528 Demecarium bromide 7.4875 Dequalinium chloride 7.3513 Piperonyl butoxide 7.4850 Adrenalone hydrochloride 7.3391 Tranilast 7.4830 Cefminox Sodium 7.3256 Nimodipine 7.4707 Lafutidine 7.3195 Procainamide hydrochloride 7.5212 Macitentan 7.3158 Fluvastatin sodium salt 7.5208 Drofenine hydrochloride 7.3145 6 Yun Y et al. J Biomed Res, 2019, 0(0)

Supplementary Table 1 Potential drugs obtained through high-throughput screening (continued) Name Score Name Score Lumefantrine 7.5192 Azilsartan Medoxomil 7.3105 Alprenolol 7.5171 Lobeline hydrochloride 7.2963 Glafenine 7.5130 Procaterol hydrochloride 7.2944 Sulfadimethoxine 7.5038 ABT530 7.2939 Sodium cromoglycate 7.4999 Cilostazo 7.2857 Demecarium bromide 7.4875 Pemetrexed acid 7.2839 Piperonyl butoxide 7.4850 Lovastatin 7.2627 Tranilast 7.4830 Erdafitinib 7.2585 Beta-Carotene 7.2469 Torezolid 7.1083 Indacaterol maleate 7.2405 Sunitinib 7.1049 Revefenacin 7.2374 Eliglustat 7.0971 Atosiban acetate 7.2316 Paromomycin Sulfate 7.0955 Candesartan Cilexetil 7.2311 Metoclopramide hydrochloride 7.0907 Cefpiramide acid 7.2226 Tannic acid 7.0870 Verteporfin 7.2222 Ritodrine hydrochloride 7.0813 Troxerutin 7.2038 Hydroxychloroquine sulfate 7.0813 Pemetrexed Disodium Hydrate 7.1984 Cimetidine 7.0729 Acecainide 7.1941 ABT199 7.0678 Phthalylsulfacetamide 7.1843 Xylitol 7.0648 Neratinib 7.1802 Safinamide mesylate 7.0647 Oxaprozin 7.1727 Pacritinib 7.0602 Sparfloxacin 7.1694 Telmisartan 7.0599 Propitocaine hydrochloride 7.1635 Nebivolol hydrochloride 7.0593 Jaceosidin 7.1613 Siponimod 7.0582 Carazolol 7.1592 Teniposide 7.0546 Ramatroban 7.1567 Benazepril hydrochloride 7.0539 Sodium gluconate 7.1536 Eletriptan HBr 7.0487 Atracurium besylate 7.1484 Methocarbamol 7.0482 Valbenazine 7.1457 Pravastatin sodium 7.0438 Tilorone dihydrochloride 7.1447 Vemurafenib 7.0314 Treprostinil Sodium 7.1396 Mycophenolate Mofetil 7.0290 Formoterol 7.1373 Imatinib Mesylate 7.0231 Cefdinir 7.1274 Lubiprostone 7.0227 Brinzolamide 7.1264 Azelaic acid 7.0209 Brexpiprazole 7.1233 Olodaterol 7.0168 Vitamin E Acetate 7.1201 Ebastine 7.0052 Fenofibrate 7.1185 Ospemifene 7.0027

a.Antiviral drugs. b.Has not been approved by FDA and was screened by molecular docking.