WO2022166885A1 - Recombinant super-compound interferon (rsifn-co) for treating covid-19 patients with or without symptoms - Google Patents
Recombinant super-compound interferon (rsifn-co) for treating covid-19 patients with or without symptoms Download PDFInfo
- Publication number
- WO2022166885A1 WO2022166885A1 PCT/CN2022/074977 CN2022074977W WO2022166885A1 WO 2022166885 A1 WO2022166885 A1 WO 2022166885A1 CN 2022074977 W CN2022074977 W CN 2022074977W WO 2022166885 A1 WO2022166885 A1 WO 2022166885A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- rsifn
- covid
- subject
- spray
- days
- Prior art date
Links
- 208000025721 COVID-19 Diseases 0.000 title claims abstract description 122
- 102000014150 Interferons Human genes 0.000 title claims abstract description 59
- 108010050904 Interferons Proteins 0.000 title claims abstract description 59
- 229940079322 interferon Drugs 0.000 title claims abstract description 53
- 208000024891 symptom Diseases 0.000 title description 40
- 239000007921 spray Substances 0.000 claims abstract description 39
- 238000000034 method Methods 0.000 claims abstract description 38
- 210000003800 pharynx Anatomy 0.000 claims abstract description 16
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 claims abstract description 15
- 229940097496 nasal spray Drugs 0.000 claims abstract description 11
- 239000007922 nasal spray Substances 0.000 claims abstract description 11
- 208000001528 Coronaviridae Infections Diseases 0.000 claims abstract description 8
- 238000010253 intravenous injection Methods 0.000 claims abstract description 8
- 238000011282 treatment Methods 0.000 claims description 34
- 201000010099 disease Diseases 0.000 claims description 32
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 32
- 208000031504 Asymptomatic Infections Diseases 0.000 claims description 21
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- 108090000623 proteins and genes Proteins 0.000 claims description 11
- 239000013078 crystal Substances 0.000 claims description 10
- 239000000203 mixture Substances 0.000 claims description 10
- 102000004169 proteins and genes Human genes 0.000 claims description 10
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical group OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 claims description 9
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid group Chemical group C(CC(O)(C(=O)O)CC(=O)O)(=O)O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 9
- 230000000694 effects Effects 0.000 claims description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N glycerol group Chemical group OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 9
- RWWYLEGWBNMMLJ-MEUHYHILSA-N remdesivir Drugs C([C@@H]1[C@H]([C@@H](O)[C@@](C#N)(O1)C=1N2N=CN=C(N)C2=CC=1)O)OP(=O)(N[C@@H](C)C(=O)OCC(CC)CC)OC1=CC=CC=C1 RWWYLEGWBNMMLJ-MEUHYHILSA-N 0.000 claims description 8
- RWWYLEGWBNMMLJ-YSOARWBDSA-N remdesivir Chemical compound NC1=NC=NN2C1=CC=C2[C@]1([C@@H]([C@@H]([C@H](O1)CO[P@](=O)(OC1=CC=CC=C1)N[C@H](C(=O)OCC(CC)CC)C)O)O)C#N RWWYLEGWBNMMLJ-YSOARWBDSA-N 0.000 claims description 8
- 241000711573 Coronaviridae Species 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical group [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 239000006172 buffering agent Substances 0.000 claims description 4
- 150000004696 coordination complex Chemical class 0.000 claims description 4
- 210000002216 heart Anatomy 0.000 claims description 4
- 239000003381 stabilizer Substances 0.000 claims description 4
- 239000004094 surface-active agent Substances 0.000 claims description 4
- 239000004034 viscosity adjusting agent Substances 0.000 claims description 4
- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical group [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 claims description 3
- 108091006905 Human Serum Albumin Proteins 0.000 claims description 3
- 102000008100 Human Serum Albumin Human genes 0.000 claims description 3
- 235000019445 benzyl alcohol Nutrition 0.000 claims description 3
- UREBDLICKHMUKA-CXSFZGCWSA-N dexamethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(F)[C@@H]1[C@@H]1C[C@@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O UREBDLICKHMUKA-CXSFZGCWSA-N 0.000 claims description 3
- 229960003957 dexamethasone Drugs 0.000 claims description 3
- 210000002249 digestive system Anatomy 0.000 claims description 3
- XXSMGPRMXLTPCZ-UHFFFAOYSA-N hydroxychloroquine Chemical compound ClC1=CC=C2C(NC(C)CCCN(CCO)CC)=CC=NC2=C1 XXSMGPRMXLTPCZ-UHFFFAOYSA-N 0.000 claims description 3
- 229960004171 hydroxychloroquine Drugs 0.000 claims description 3
- 210000003734 kidney Anatomy 0.000 claims description 3
- 210000004185 liver Anatomy 0.000 claims description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 3
- 229920000053 polysorbate 80 Polymers 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- -1 bacteriostat Substances 0.000 claims description 2
- 238000012360 testing method Methods 0.000 description 33
- 239000003814 drug Substances 0.000 description 26
- 210000004027 cell Anatomy 0.000 description 21
- 229940079593 drug Drugs 0.000 description 21
- 230000006872 improvement Effects 0.000 description 18
- 210000004369 blood Anatomy 0.000 description 16
- 239000008280 blood Substances 0.000 description 16
- 238000002347 injection Methods 0.000 description 16
- 239000007924 injection Substances 0.000 description 16
- 239000000825 pharmaceutical preparation Substances 0.000 description 16
- 239000000047 product Substances 0.000 description 16
- 239000000523 sample Substances 0.000 description 16
- 241001678559 COVID-19 virus Species 0.000 description 15
- 229940126534 drug product Drugs 0.000 description 15
- 108010079944 Interferon-alpha2b Proteins 0.000 description 14
- 229940068196 placebo Drugs 0.000 description 14
- 239000000902 placebo Substances 0.000 description 14
- 108020000999 Viral RNA Proteins 0.000 description 13
- 102100040018 Interferon alpha-2 Human genes 0.000 description 12
- 238000002565 electrocardiography Methods 0.000 description 12
- 238000012216 screening Methods 0.000 description 12
- 241000700605 Viruses Species 0.000 description 11
- 238000002483 medication Methods 0.000 description 11
- 210000002345 respiratory system Anatomy 0.000 description 11
- 150000001413 amino acids Chemical group 0.000 description 10
- 241000589516 Pseudomonas Species 0.000 description 9
- 206010037660 Pyrexia Diseases 0.000 description 9
- 238000013381 RNA quantification Methods 0.000 description 9
- 238000003384 imaging method Methods 0.000 description 9
- 238000012797 qualification Methods 0.000 description 9
- 208000000059 Dyspnea Diseases 0.000 description 8
- 206010013975 Dyspnoeas Diseases 0.000 description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 8
- 229910052760 oxygen Inorganic materials 0.000 description 8
- 239000001301 oxygen Substances 0.000 description 8
- 231100001274 therapeutic index Toxicity 0.000 description 8
- 230000003612 virological effect Effects 0.000 description 8
- 239000008186 active pharmaceutical agent Substances 0.000 description 7
- 210000000038 chest Anatomy 0.000 description 7
- 208000015181 infectious disease Diseases 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 206010011224 Cough Diseases 0.000 description 6
- 206010019233 Headaches Diseases 0.000 description 6
- 208000000112 Myalgia Diseases 0.000 description 6
- 206010035664 Pneumonia Diseases 0.000 description 6
- 238000010240 RT-PCR analysis Methods 0.000 description 6
- 231100000869 headache Toxicity 0.000 description 6
- 229940047124 interferons Drugs 0.000 description 6
- 238000009533 lab test Methods 0.000 description 6
- 238000002663 nebulization Methods 0.000 description 6
- 150000007523 nucleic acids Chemical class 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 229920002477 rna polymer Polymers 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 208000010470 Ageusia Diseases 0.000 description 5
- 206010002653 Anosmia Diseases 0.000 description 5
- 206010068319 Oropharyngeal pain Diseases 0.000 description 5
- 201000007100 Pharyngitis Diseases 0.000 description 5
- 208000037847 SARS-CoV-2-infection Diseases 0.000 description 5
- 230000002411 adverse Effects 0.000 description 5
- 235000019666 ageusia Nutrition 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000003013 cytotoxicity Effects 0.000 description 5
- 231100000135 cytotoxicity Toxicity 0.000 description 5
- 229940088679 drug related substance Drugs 0.000 description 5
- 238000004519 manufacturing process Methods 0.000 description 5
- 108020004707 nucleic acids Proteins 0.000 description 5
- 102000039446 nucleic acids Human genes 0.000 description 5
- 230000001681 protective effect Effects 0.000 description 5
- 238000003753 real-time PCR Methods 0.000 description 5
- 238000012552 review Methods 0.000 description 5
- 208000013220 shortness of breath Diseases 0.000 description 5
- 238000010561 standard procedure Methods 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 230000002159 abnormal effect Effects 0.000 description 4
- 239000000427 antigen Substances 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 238000010609 cell counting kit-8 assay Methods 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 206010016256 fatigue Diseases 0.000 description 4
- 238000003306 harvesting Methods 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 239000013612 plasmid Substances 0.000 description 4
- 230000004224 protection Effects 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 108010014726 Interferon Type I Proteins 0.000 description 3
- 102000002227 Interferon Type I Human genes 0.000 description 3
- 206010028735 Nasal congestion Diseases 0.000 description 3
- 208000004756 Respiratory Insufficiency Diseases 0.000 description 3
- 206010040070 Septic Shock Diseases 0.000 description 3
- 208000036142 Viral infection Diseases 0.000 description 3
- 208000022531 anorexia Diseases 0.000 description 3
- 235000019558 anosmia Nutrition 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 3
- 206010061428 decreased appetite Diseases 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 208000002672 hepatitis B Diseases 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 210000004072 lung Anatomy 0.000 description 3
- 238000013411 master cell bank Methods 0.000 description 3
- 229910021645 metal ion Inorganic materials 0.000 description 3
- 230000008383 multiple organ dysfunction Effects 0.000 description 3
- DILRJUIACXKSQE-UHFFFAOYSA-N n',n'-dimethylethane-1,2-diamine Chemical compound CN(C)CCN DILRJUIACXKSQE-UHFFFAOYSA-N 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 238000003752 polymerase chain reaction Methods 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 230000000241 respiratory effect Effects 0.000 description 3
- 201000004193 respiratory failure Diseases 0.000 description 3
- 230000036391 respiratory frequency Effects 0.000 description 3
- 230000036303 septic shock Effects 0.000 description 3
- 230000009385 viral infection Effects 0.000 description 3
- 102100035765 Angiotensin-converting enzyme 2 Human genes 0.000 description 2
- 108090000975 Angiotensin-converting enzyme 2 Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 241000700721 Hepatitis B virus Species 0.000 description 2
- 101000959820 Homo sapiens Interferon alpha-1/13 Proteins 0.000 description 2
- 206010021143 Hypoxia Diseases 0.000 description 2
- 102100040019 Interferon alpha-1/13 Human genes 0.000 description 2
- 108010047761 Interferon-alpha Proteins 0.000 description 2
- 102000006992 Interferon-alpha Human genes 0.000 description 2
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 2
- 102000003960 Ligases Human genes 0.000 description 2
- 108090000364 Ligases Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 206010035737 Pneumonia viral Diseases 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- SIIZPVYVXNXXQG-KGXOGWRBSA-N [(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-4-[[(3s,4r)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-3-hydroxyoxolan-2-yl]methyl [(2r,4r,5r)-2-(6-aminopurin-9-yl)-4-hydroxy-5-(phosphonooxymethyl)oxolan-3-yl] hydrogen phosphate Polymers C1=NC2=C(N)N=CN=C2N1[C@@H]1O[C@H](COP(O)(=O)OC2[C@@H](O[C@H](COP(O)(O)=O)[C@H]2O)N2C3=NC=NC(N)=C3N=C2)[C@@H](O)[C@H]1OP(O)(=O)OCC([C@@H](O)[C@H]1O)OC1N1C(N=CN=C2N)=C2N=C1 SIIZPVYVXNXXQG-KGXOGWRBSA-N 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 229910002091 carbon monoxide Inorganic materials 0.000 description 2
- 208000020832 chronic kidney disease Diseases 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000035487 diastolic blood pressure Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 229920001903 high density polyethylene Polymers 0.000 description 2
- 239000004700 high-density polyethylene Substances 0.000 description 2
- 230000007954 hypoxia Effects 0.000 description 2
- 230000002519 immonomodulatory effect Effects 0.000 description 2
- 210000003000 inclusion body Anatomy 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 108010010648 interferon alfacon-1 Proteins 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 206010025482 malaise Diseases 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 208000013465 muscle pain Diseases 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 238000009597 pregnancy test Methods 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 208000023504 respiratory system disease Diseases 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 230000003319 supportive effect Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- KCFYEAOKVJSACF-UHFFFAOYSA-N umifenovir Chemical compound CN1C2=CC(Br)=C(O)C(CN(C)C)=C2C(C(=O)OCC)=C1CSC1=CC=CC=C1 KCFYEAOKVJSACF-UHFFFAOYSA-N 0.000 description 2
- 229960004626 umifenovir Drugs 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 230000029812 viral genome replication Effects 0.000 description 2
- 208000009421 viral pneumonia Diseases 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 239000008215 water for injection Substances 0.000 description 2
- 102100027962 2-5A-dependent ribonuclease Human genes 0.000 description 1
- 108010000834 2-5A-dependent ribonuclease Proteins 0.000 description 1
- ZOOGRGPOEVQQDX-UUOKFMHZSA-N 3',5'-cyclic GMP Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=C(NC2=O)N)=C2N=C1 ZOOGRGPOEVQQDX-UUOKFMHZSA-N 0.000 description 1
- 101710169336 5'-deoxyadenosine deaminase Proteins 0.000 description 1
- 102100036664 Adenosine deaminase Human genes 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 208000020446 Cardiac disease Diseases 0.000 description 1
- 108010001857 Cell Surface Receptors Proteins 0.000 description 1
- 208000000419 Chronic Hepatitis B Diseases 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 108090000204 Dipeptidase 1 Proteins 0.000 description 1
- 206010061818 Disease progression Diseases 0.000 description 1
- 108091029865 Exogenous DNA Proteins 0.000 description 1
- 241001069765 Fridericia <angiosperm> Species 0.000 description 1
- 101000852870 Homo sapiens Interferon alpha/beta receptor 1 Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 206010062016 Immunosuppression Diseases 0.000 description 1
- 206010022004 Influenza like illness Diseases 0.000 description 1
- 102100036714 Interferon alpha/beta receptor 1 Human genes 0.000 description 1
- 102100026720 Interferon beta Human genes 0.000 description 1
- 108090000467 Interferon-beta Proteins 0.000 description 1
- KJHKTHWMRKYKJE-SUGCFTRWSA-N Kaletra Chemical compound N1([C@@H](C(C)C)C(=O)N[C@H](C[C@H](O)[C@H](CC=2C=CC=CC=2)NC(=O)COC=2C(=CC=CC=2C)C)CC=2C=CC=CC=2)CCCNC1=O KJHKTHWMRKYKJE-SUGCFTRWSA-N 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 208000019693 Lung disease Diseases 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000204031 Mycoplasma Species 0.000 description 1
- 208000031662 Noncommunicable disease Diseases 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 208000003251 Pruritus Diseases 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- NCDNCNXCDXHOMX-UHFFFAOYSA-N Ritonavir Natural products C=1C=CC=CC=1CC(NC(=O)OCC=1SC=NC=1)C(O)CC(CC=1C=CC=CC=1)NC(=O)C(C(C)C)NC(=O)N(C)CC1=CSC(C(C)C)=N1 NCDNCNXCDXHOMX-UHFFFAOYSA-N 0.000 description 1
- 241000315672 SARS coronavirus Species 0.000 description 1
- 241001678558 SARS-like-CoVZXC21 Species 0.000 description 1
- 108700009124 Transcription Initiation Site Proteins 0.000 description 1
- 208000018756 Variant Creutzfeldt-Jakob disease Diseases 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 108700003859 araC Genes Proteins 0.000 description 1
- 101150044616 araC gene Proteins 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 102000006635 beta-lactamase Human genes 0.000 description 1
- 102000023732 binding proteins Human genes 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 101150049515 bla gene Proteins 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 229940000031 blood and blood forming organ drug Drugs 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 208000005881 bovine spongiform encephalopathy Diseases 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000011976 chest X-ray Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000000978 circular dichroism spectroscopy Methods 0.000 description 1
- 235000015165 citric acid Nutrition 0.000 description 1
- 229940126523 co-drug Drugs 0.000 description 1
- 238000011278 co-treatment Methods 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000002591 computed tomography Methods 0.000 description 1
- 238000005094 computer simulation Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000000120 cytopathologic effect Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 231100000517 death Toxicity 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 231100000371 dose-limiting toxicity Toxicity 0.000 description 1
- 238000011143 downstream manufacturing Methods 0.000 description 1
- 239000006196 drop Substances 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 210000005069 ears Anatomy 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000001508 eye Anatomy 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 210000001061 forehead Anatomy 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 210000005095 gastrointestinal system Anatomy 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 229940090438 infergen Drugs 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 230000028709 inflammatory response Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 229960003358 interferon alfacon-1 Drugs 0.000 description 1
- 230000011488 interferon-alpha production Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 230000007803 itching Effects 0.000 description 1
- 229960004525 lopinavir Drugs 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 238000001840 matrix-assisted laser desorption--ionisation time-of-flight mass spectrometry Methods 0.000 description 1
- 238000005399 mechanical ventilation Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 102000006240 membrane receptors Human genes 0.000 description 1
- 230000003340 mental effect Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 210000002346 musculoskeletal system Anatomy 0.000 description 1
- OHDXDNUPVVYWOV-UHFFFAOYSA-N n-methyl-1-(2-naphthalen-1-ylsulfanylphenyl)methanamine Chemical compound CNCC1=CC=CC=C1SC1=CC=CC2=CC=CC=C12 OHDXDNUPVVYWOV-UHFFFAOYSA-N 0.000 description 1
- 239000007923 nasal drop Substances 0.000 description 1
- 229940100662 nasal drops Drugs 0.000 description 1
- 210000002850 nasal mucosa Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 210000001331 nose Anatomy 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 description 1
- 229940068968 polysorbate 80 Drugs 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000009979 protective mechanism Effects 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 238000012429 release testing Methods 0.000 description 1
- 238000004153 renaturation Methods 0.000 description 1
- 230000036387 respiratory rate Effects 0.000 description 1
- 230000000284 resting effect Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- NCDNCNXCDXHOMX-XGKFQTDJSA-N ritonavir Chemical compound N([C@@H](C(C)C)C(=O)N[C@H](C[C@H](O)[C@H](CC=1C=CC=CC=1)NC(=O)OCC=1SC=NC=1)CC=1C=CC=CC=1)C(=O)N(C)CC1=CSC(C(C)C)=N1 NCDNCNXCDXHOMX-XGKFQTDJSA-N 0.000 description 1
- 229960000311 ritonavir Drugs 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000011265 semifinished product Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 235000002639 sodium chloride Nutrition 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 230000000153 supplemental effect Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000035488 systolic blood pressure Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000005030 transcription termination Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 230000007485 viral shedding Effects 0.000 description 1
- 238000009528 vital sign measurement Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
- A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
- A61K31/404—Indoles, e.g. pindolol
- A61K31/4045—Indole-alkylamines; Amides thereof, e.g. serotonin, melatonin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/425—Thiazoles
- A61K31/427—Thiazoles not condensed and containing further heterocyclic rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4706—4-Aminoquinolines; 8-Aminoquinolines, e.g. chloroquine, primaquine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/513—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim having oxo groups directly attached to the heterocyclic ring, e.g. cytosine
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
- A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/21—Interferons [IFN]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
Definitions
- the present invention relates to recombinant super-compound interferon (rSIFN-co) and its use to treat COVID-19 patients with or without symptoms.
- COVID-19 Common symptoms of COVID-19 include fever (83–99%) , cough (59–82%) , fatigue (44–70%) , anorexia (40–84%) , shortness of breath (31–40%) , and myalgias (11–35%) [3] . Other reported symptoms have included, but are not limited to, sore throat, nasal congestion, headache, diarrhea, nausea and vomiting, anosmia, and ageusia [3] . The mortality of COVID-19 varies by geographical area. According to a recent analysis conducted by the Centers for Disease Control and Prevention (CDC) of the United States, more than 1.3 million laboratory-confirmed cases were reported between January and May 2020.
- CDC Centers for Disease Control and Prevention
- COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) , a newly discovered virus in the family of Coronaviridae (beta) , which has 89%nucleotide identity with bat SARS-like-CoVZXC21 and 82%with human SARS-CoV [6] .
- SARS-CoV-2 is an enveloped, positive-sense, single-stranded RNA virus. Virus entry is achieved through binding of viral spike (S) protein to its receptor, angiotensin-converting enzyme 2 (ACE2) , on the host cell [7] .
- SARS-CoV-2 severe acute respiratory syndrome coronavirus 2
- ACE2 angiotensin-converting enzyme 2
- the present invention discloses a method that prevents and/or treats COVID-19 (SARS-Cov-2) coronavirus infection, including mild COVID-19 infection and asymptomatic infection, in a subject suffering from diseases in digestive system, liver, heart and/or kidney.
- the method comprises administering to the subject therapeutically effective doses, e.g., from 5 to 100 ⁇ g per day, of recombinant super-compound interferon (rSIFN-co) for a period of at least 3 days and up to 30 days.
- the administration can be by nasal spray or intravenous injection.
- Figure 1 is an illustration of the study schema.
- Figure 2 compares structure of rSIFN-co and IFN- ⁇ 2b
- Figure 3 compares structure of rSIFN-co and Infergen of Amgen
- Figure 4 shows vector diagram of rSIFN-co in Plasmid pHY-5.
- Figure 5 shows a manufacturing flowchart for rSIFN-co drug product.
- interferons have long been used as antiviral treatments.
- the preclinical studies of rSIFN-co have demonstrated its superiority of anti-SARS-CoV-2 capacity than conventional IFNs.
- dysregulated IFNs was found in COVID-19 patients, implicating the role of IFN in COVID-19 pathogenesis [18] .
- the potential of rSIFN-co as a protective and therapeutic agent against SARS-CoV-2 infection is suggested.
- Impairment of type I interferon (IFN) response was discovered in severe and critical COVID-19 patients. No IFN- ⁇ and low IFN- ⁇ production/activity were observed and was associated with a persistent blood viral load and an exacerbated inflammatory response [17] .
- IFNs type I interferon
- IFN ⁇ -1a the efficacy and safety of IFN ⁇ -1a were evaluated in patients with severe COVID-19.
- the patients in the IFN group received subcutaneous injection of 12 million IU/mL of IFN ⁇ -1a three times per week for two weeks [22] .
- time to reach clinical response was not significantly improved, the proportions of subjects discharged on Day 14 were significantly higher in the IFN group [22] .
- Recombinant super-compound interferon is a product of patented technological research developed by Sichuan Huiyang Life Science and Technology Corporation. It is a recombinant form of the naturally occurring cytokine interferon-alpha (IFN- ⁇ ) which has a modified spatial configuration.
- rSIFN-co has the same amino acid sequence as the Amgen product (Interferon Alfacon-1) consisting of 167 amino acids (MCDLPQTHSLGNRRALILLAQMRRISPFSCLKDRHDFGFPQEEFDGNQFQKAQAISVLHEMIQQTFNLFSTKDSSAAWDESLLEKFYTELYQQLNDLEACVIQEVGVEETPLMNVDSILAVKKYFQRITLYLTEKKYSPCAWEVVRAEIMRSFSLSTNLQERLRRKE, SEQ ID NO: 1) .
- the amino acid homology of and rSIFN-co are naturally occurring interferon ⁇ -2a and interferon ⁇ -2b, with only a difference of 18 and 19 amino acids, respectively.
- rSIFN-co This amino acid difference results in rSIFN-co having 10 times greater affinity for IFN-specific cell surface receptors IFNAR1 compared to interferon ⁇ -2a and interferon ⁇ -2b [15] .
- rSIFN-co binds to IFN-specific cell surface receptors, resulting in the transcription and translation of genes whose protein products have antiviral, antiproliferative, anticancer, and immune-modulating effects.
- the 3-dimensional conformational change improves efficacy and causes fewer side effects compared to IFN-a.
- the nucleic acid sequence encoding the amino acid sequence of SEQ ID NO. 1 is as follows (SEQ ID NO: 2) .
- the rSIFN-co can be produced by the method comprising the following steps: introducing a nucleotide sequence comprising SEQ ID NO: 2 that encodes the recombinant interferon into an isolated host cell; culturing the host cell under appropriate condition for expression of the recombinant interferon; and harvesting the recombinant interferon, wherein the recombinant interferon has an amino acid sequence of SEQ ID NO: 1, and the recombinant interferon inhibits secretion of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) of Hepatitis B Virus.
- the host cell is Escherichia coli.
- the nucleotide sequence comprising SEQ ID NO: 2 is under the control of the promoter PBAD.
- the harvesting step comprises extraction of the interferon from the fermentation broth, collection of the inclusion bodies, denaturation and renaturation of the harvested interferon. Still further, the harvesting step also comprises separation and purification of the recombinant interferon. Met at the N-terminus of SEQ ID NO: 1 is resected in mature protein.
- the study used reagents including DMEM (Gibco) , fetal calf serum (Gibco) , DMSO (sigma) , double-antibody, pancreatine, etc., kits including Cell Counting Kit-8 (CCK-8) (B34304, bimake) , QIAamp viral RNA mini kit (52906, Qiagen) , Novel Coronavirus (2019-nCoV) Real Time RT-PCR kit (by Wuhan Institute of Virology) , and main instruments such as Varioskan Flash (by Thermo Fisher Scientific) , QIAcube HT 9001793 (by Qiagen) and CFX96 Touch Real-Time PCR Detection System (by Bio-rad) .
- DMEM Gibco
- Gibco fetal calf serum
- DMSO double-antibody
- pancreatine pancreatine, etc.
- kits including Cell Counting Kit-8 (CCK-8) (B34304, bimake)
- the final set of concentrations of rSIFN-co was 6 ⁇ 10 8 , 3 ⁇ 10 8 , 1.5 ⁇ 10 8 , 7.5 ⁇ 10 7 , 3.75 ⁇ 10 7 , 1.875 ⁇ 10 7 and 0 pg/ml
- the final set of concentrations of recombinant human Interferon ⁇ -2b injection was 2 ⁇ 10 7 , 1 ⁇ 10 7 , 5 ⁇ 10 6 , 2.5 ⁇ 10 6 , 1.25 ⁇ 10 6 , 6.25 ⁇ 10 5 and 0 pg/ml
- the final set of concentrations of Remdesivir was 320, 160, 80, 40, 20, 10 and 0 ⁇ M;
- a) Grow Vero-E6 cells in a 24-well plate, 8 ⁇ 10 4 cells per well. Put the plate into 37 °Cincubator with 5%CO 2 . When the confluence reaches 70%-80%, using DMEN with 2%FBS for 2 times gradient dilution of rSIFN-co, recombinant human Interferon ⁇ -2b injection (pseudomonas) and Remdesivir, respectively.
- qRT-PCR real time RT-PCR
- RNA quantitative detection on collected virus. Take 20 ⁇ L collected supernatant after infection and extract RNA following instructions of QIAamp viral RNA mini kit. Conduct the qRT-PCR by using Novel Coronavirus (2019-nCoV) Real Time RT-PCR kit (TaqMan probe method) ;
- Inhibition rate (%) 1 -Viral RNA Copies in test groups /Viral RNA Copies in drug free group ⁇ 100%. Analyze the half effective concentration of drugs (EC 50 ) by GraphPad PrisM6.0; and
- Therapeutic Index (TI) half cytotoxicity concentration (CC 50 ) /half effective concentration (EC 50 ) .
- the study showed that the EC 50 and CC 50 of rSIFN-co were 14.6 pg/mL and 7.12 ⁇ 10 8 pg/mL, respectively, whereas the EC 50 and CC 50 of IFN ⁇ -2b were 162 pg/mL and 1.37 ⁇ 10 7 pg/mL, respectively.
- the results suggested that rSIFN-co exhibited a higher efficacy against SARS-CoV-2 and a lower cytotoxicity as compared with IFN ⁇ -2b.
- PK pharmacokinetic
- rSIFN-co induces more 2′-5′-oligoadenylate synthetase (2′, 5′-OAS; a specific marker triggered by interferon) over a longer time period, when compared to 2′, 5′-OAS induced by
- the incidence of adverse events in the rSIFN-co group was lower than that in the group.
- Type I interferons are known to play an important role in first-line defense against viruses by stimulating the expression of proteins such as ribonucleic acid (RNA) -dependent protein kinase, 2′, 5′-OAS, RNase L, and RNA-specific adenosine deaminase [16] .
- RNA ribonucleic acid
- 2′, 5′-OAS ribonucleic acid
- RNase L RNA-specific adenosine deaminase
- rSIFN-co severe acute respiratory syndrome
- Sichuan aprovince in China
- rSIFN-co spray was allocated to 3,000 users, including doctors and nurses in hospitals, populated areas with a high risk for SARS, and the National research group.
- the highest dose was 16 million international unit (IU) and the longest treatment duration was over two months. No side effects connected to the use of the spray was reported, and none of users administered with rSIFN-co spray has been infected by SARS.
- the present invention provides a method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject.
- the method comprises administering to the subject a therapeutically effective dose of recombinant super-compound interferon (rSIFN-co) for a period of at least 3 days.
- rSIFN-co recombinant super-compound interferon
- the rSIFN-co is administered to said subject by nasal spray, pharynx spray or intravenous injection.
- the therapeutically effective dose ranges from 5 ⁇ g to 100 ⁇ g per day.
- the therapeutically effective dose is selected from the group consisting of 8 ⁇ g, 16 ⁇ g, 32 ⁇ g, 64 ⁇ g and 96 ⁇ g per day.
- the period is 5 to 30 days.
- the subject suffers from one or more diseases in one or more of the digestive system, liver, heart, and kidney.
- the subject receives one or more additional treatments selected from the group consisting of Remdesivir, hydroxychloroquine, and dexamethasone.
- the subject is healthy or has an asymptomatic infection of Covid-19 coronavirus, and said subject receives 2 sprays per nostril and 4 sprays through pharynx once daily, each spray comprising 0.2-5.0 ⁇ g of said rSIFN-co.
- the subject shows mild covid-19 infection and receives 2 sprays per nostril and 4 sprays through pharynx twice daily, each spray comprising 0.2-5.0 ⁇ g of said rSIFN-co.
- the rSIFN-co has a specific activity greater than 5.0 x 10 8 IU/mg protein.
- the nasal spray, pharynx spray or intravenous injection comprises one or more of stabilizer, bacteriostat, surfactant, metal complex, isosmotic adjusting agent, buffering agent, and viscosity modifier.
- the stabilizer is human serum albumin.
- the metal complex is disodium edetate.
- the surfactant is Tween 80.
- the buffering agent is citric acid.
- the viscosity modifier is glycerol.
- the isosmotic adjusting agent is sodium chloride.
- the bacteriostat is benzyl alcohol.
- the nasal spray, pharynx spray or intravenous injection has a pH value of about 5.0.
- the nasal spray or pharynx spray is stable under 4-8°C for 3 months, at 25°C for 2 months and at 37°C for 2 weeks.
- the present provides a method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject.
- the method comprises administering to the subject a composition comprising a therapeutically effective dose of recombinant super-compound interferon (rSIFN-co) for a period of at least 3 days
- the present provides a method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject by administering a composition consisting of a therapeutically effective dose of rSIFN-co for a period of at least 3 days.
- SARS-Cov-2 Covid-19 coronavirus
- the primary objective is to assess the safety and tolerability of rSIFN-co in healthy subjects in close contact with confirmed COVID-19 case (s) and patients with mild COVID-19 or asymptomatic infection.
- the secondary objective is to assess the efficacy of rSIFN-co in healthy subjects in close contact with confirmed COVID-19 case (s) and patients with mild COVID-19 or asymptomatic infection.
- the primary endpoint is the safety and tolerability profiles of rSIFN-co.
- the secondary endpoint for Healthy subjects in close contact with confirmed COVID-19 case (s) is:
- the secondary endpoint for patients with mild COVID-19 or asymptomatic infection is:
- the exploratory endpoint for Healthy subjects in close contact with confirmed COVID-19 case (s) is the subjects’ evaluation on ease of use, discomfort using the study product, and overall satisfaction.
- the exploratory endpoint for patients with mild COVID-19 or asymptomatic infection is:
- the study is designed as a randomized, double-blind, placebo-controlled study to minimize the potential for subjective bias.
- the inclusion of placebo arms will provide a control for comparison of efficacy endpoints between the treatment groups.
- the study will be conducted at multiple study centers to facilitate subject enrollment and to increase the generalizability of the study results.
- the doses evaluated for this study 8 and 16 million IU, are based on the totality of the data from previous trials in adults.
- An eligible subject must fulfill ALL of the following inclusion criteria:
- RT-PCR Negative reverse transcriptase polymerase chain reaction
- Healthy subjects in close contact with confirmed COVID-19 case (s) Subjects with exposure to confirmed COVID-19 case (s) within 96 hours and belong to one of the following exposure situations per health department directives:
- ICU intensive care unit
- First responders i.e., first aid personnel, paramedics
- Level 2 protection refers to wearing a disposable cap, medical protective mask (N95 or higher level medical protective mask) , goggles (anti-fog type) or protective mask (anti-fog type) , medical protective suit or overalls (white gown) , disposable gloves and disposable shoe covers.
- Asymptomatic or presymptomatic infection Individuals who test positive for SARS-CoV-2 by virologic testing using a molecular diagnostic (e.g., polymerase chain reaction) or antigen test, but have no symptoms.
- a molecular diagnostic e.g., polymerase chain reaction
- antigen test e.g., antigen test, but have no symptoms.
- rSIFN-co or placebo, for a maximum of 10 to 28 days.
- rSIFN-co will be administered with one or more other agents, wherein the control group will receive said one or more other agents only.
- rSIFN-co The dose of rSIFN-co given by nebulization or injection is equivalent to 8 to 16 million IU (32 ⁇ g) per day. Other agents will be given in their usual dosages.
- Randomize eligible subjects into one of the two treatment arms i.e., placebo or rSIFN-co
- Vital sign measurements include blood pressures (systolic blood pressure [SBP] and diastolic blood pressure [DBP] ) , pulse rate, respiratory rate, body temperature, and SpO 2 . Vital signs will be assessed at each visit. Results will be documented in the CRF. Measurements of all vital signs shall be completed in the during the visit within a time frame of 2 hours. Measurements should be taken in a seated position after resting for approximately 10 minutes. The same thermometer should be used consistently throughout the study. Subjects in Group B will be provided with an oximeter to help monitor their oxygen level. The subjects will be instructed to document daily oxygen level on the diary card.
- SBP systolic blood pressure
- DBP diastolic blood pressure
- Fever is defined as rectal, ear, or forehead temperature greater than 38 °C. Site personnel should measure ear temperature of subject if possible. If other methods are used, the same method should be used consistently throughout the study.
- a complete physical examination will be conducted by the Investigator or designee at screening.
- Complete physical examination items include general appearance, HEENT (head, eyes, ears, nose, and throat) , mouth, skin, neck (including thyroid) , lymph nodes, spine, cardiovascular system, respiratory system, gastro-intestinal system, nervous system, musculoskeletal system, blood and blood forming organs, mental status, and other body systems if applicable for describing the status of subject’s health.
- HEENT head, eyes, ears, nose, and throat
- mouth skin, neck (including thyroid)
- lymph nodes spine
- cardiovascular system respiratory system
- gastro-intestinal system gastro-intestinal system
- nervous system musculoskeletal system
- blood and blood forming organs forming organs, mental status, and other body systems if applicable for describing the status of subject’s health.
- an abbreviated physical examination comprising a brief medical history and targeted physical examination will be performed at the discretion of the Investigator to evaluate possible adverse events.
- Electrocardiogram ECG
- Single 12-lead ECG will be obtained at screening using an ECG machine that automatically calculates the heart rate and measures PR, QRS, QT, and QTc (either Bazette or Fridericia formulas) intervals.
- the assessment will follow the current SOPs at each participating study center. Throughout the study, additional ECGs may be obtained if clinically indicated.
- the Investigators will compare ECGs to the screening ECG and document overall interpretation of specific ECGs as either normal, abnormal with clinical significance, or abnormal without clinical significance. An AE will be recorded if clinically significant.
- Laboratory tests (including hematology, biochemistry, and coagulation) will be performed at screening, Days 5, 10, 15, 21, 28, and 60. All samples will be analyzed by the study center’s local laboratory per the standard operating procedures (SOPs) of the laboratory.
- the examination items will include analytes in the following table:
- Additional tests may be conducted at any time during the study as determined necessary by the Investigator. A clinically significant change from baseline will be recorded as an AE.
- nasopharyngeal swabs are preferred [26] , but if these are not obtainable or additional samples are required by local regulations, oropharyngeal or nasal swabs may be obtained if necessary.
- Samples will be delivered to the laboratories selected by the Sponsor to quantitatively and qualitatively detect viral RNA via RT-PCR assays approved by local regulatory authorities. Preparation, handling, storage, shipment, and examinations of specimens will follow the lab SOPs.
- Thoracic CT-scan or chest X-ray will be performed at Screening, Day 5 (only for mild COVID-19 patients or asymptomatic infection) , and End of Treatment (Day 10) .
- a standardized framework will be used to assess the chest imaging at each study site.
- Clinical improvement will be defined as a decrease of at least one point on the 11-point scale compared to the baseline value (e.g., from 2 to 1; from 2 to 0) .
- COVID-19 disease severity and symptoms will be assessed and documented at screening and throughout the entire study. Typical symptoms for patients with mild disease include fever, cough, fatigue, anorexia, shortness of breath, myalgias, sore throat, nasal congestion, headache, gastrointestinal symptoms, loss of smell (anosmia) , loss of taste (ageusia) , and without evidence of viral pneumonia or hypoxia.
- Asymptomatic or presymptomatic infection Individuals who test positive for SARS-CoV-2 by virologic testing using a molecular diagnostic (e.g., polymerase chain reaction) or antigen test, but have no symptoms;
- Mild disease Individuals who have any of the various signs and symptoms of COVID-19 (e.g., fever, cough, sore throat, malaise, headache, muscle pain) and a SpO2 ⁇ 94%on room air at sea level, without shortness of breath, dyspnea, or abnormal chest imaging;
- COVID-19 e.g., fever, cough, sore throat, malaise, headache, muscle pain
- SpO2 ⁇ 94%on room air at sea level, without shortness of breath, dyspnea, or abnormal chest imaging
- Moderate disease Individuals who have evidence of lower respiratory disease by clinical assessment or imaging and SpO2 ⁇ 94%on room air at sea level;
- Severe disease Individuals who have respiratory frequency >30 breaths per minute, SpO2 ⁇ 94%on room air at sea level, ratio of arterial partial pressure of oxygen to fraction of inspired oxygen (PaO 2 /FiO 2 ) ⁇ 300 mmHg, or lung infiltrates >50%; and
- ⁇ Critical disease Individuals who have respiratory failure, septic shock, and/or multiple organ dysfunction.
- the Master Cell Bank (MCB) and Working Cell Bank (WCB) for rSIFN-co are manufactured under cGMP.
- the DS manufacture process involves bioproduction and harvest then purification of rSIFN-co from the E. coli inclusion bodies using upstream and downstream manufacturing processing typical for this type of production.
- the rSIFN-co cDNA was cloned into the E. coli high expression vector pHY-4 plasmid to yield the final recombinant plasmid pHY-5 ( Figure 4) .
- Typical production and purification can be found in US Patent No. 8,415,151 and Patent No. 8,846,025.
- ⁇ PBAD promoter which derived from bacterial arabinose operon (araBAD operon) and was mainly regulated by CAP-cAMP and L-arabinose binding protein AraC i.e. product of ara Cgene
- ⁇ ⁇ -Lactamase coded by Bla gene could provide ampicillin resistance.
- ⁇ ColE1 replicon determined the copy number and replication of plasmid in bacteria.
- Animal-based ingredients, tryptone and acid hydrolyzed casein, are used in upstream manufacture and are of GMP quality and certified that they are sourced in a controlled and validated way as to reduce the risk of contamination or exposure to any diseases, especially Bovine Spongiform Encephalopathy.
- the release testing specifications for the DS include protein content, biological activity, specific activity, purity, molecular weight, exogenous DNA, host protein, antibiotic residue, endotoxin and a number of characterization assays.
- the biological activity of rSIFN-co was determined based on the effect of rSIFN-co to protect cells against a viral cytopathic effect. It is quantified using a CPE potency assay, referenced against IFN ⁇ -2a reference standard, which has been tested and calibrated in International Units against the WHO reference standard.
- the three-dimensional structure of the present recombinant interferon (SEQ ID NO: 3, without N-terminal M) is different from the three-dimensional structure of IFN ⁇ -2b (SEQ ID NO: 4) published in the art (see Figure 2) and the three-dimensional structure of based on computational modeling. There are obvious differences between the AB loops of the two, and their BC loops also cannot overlap completely (see Figure 3) .
- Stability was tested according to ICH guideline for biological products Q1A and Q5C.
- the stability of the rSIFN-co composition is tested under 4-8°C for 6 months, -80°C for 24 months, and accelerated condition at 25°C for 1 month. Repeated freeze/thaw cycle tests are done for 7 cycles. Stability data for 2 representative batches of drug substance 201801003, and 201801004 is shown in the following Tables 7-10.
- a composition of rSIFN-co was prepared by obtaining a 80%of the total volume of water for injection, adding a prescribed amount of disodium edetate, polysorbate 80, citric acid, glycerol, sodium chloride and benzyl alcohol in turn while stirring. Add the next component after the former component is fully dissolved. After each component is added, stir for more than 60 minutes until well combined. Cool the solution to below 20°C and adjust pH value to about 5.0 with diluted hydrochloric acid. Stir the solution fully, add the prescribed amount of human serum albumin and rSIFN-co protein stock solution and then test pH value. If pH deviates, adjust pH value again to 5.0 and stir slightly. Dilute with water for injection cooled to below 20°C to volume, homogenizing by gently stirring for more than 30 minutes before filter and sterilize with a 0.22 ⁇ m filter to obtain the semi-finished rSIFN-co spray.
- a typical workflow is shown in Figure 5.
- composition (alternatively “drug product” ) is stable under 4-8°C for 3 months and the real time study is on-going. It is also stable under accelerated condition at 25°C for 2 months and at 37°C for 2 weeks.
- the main components of the drug product are shown in the Table 11.
- the product is a homogeneous colorless or light yellow with slightly sticky liquid spray.
- the product is filled in high-density polyethylene bottles for pharmaceuticals with a mechanical nasal spray pump. Each bottle contains 8 ml and each pump is 0.1 ml.
- a solution is prepared according to the formula followed by sterilization and filtering by a 0.22 ⁇ m filter.
- the filtered liquid was transferred into a sterile container to obtain a semi-finished product.
- the sterile filtered solution is packaged into 10 ml medicinal high-density polyethylene bottle for external use by filling machine and equip with medicinal spray pump.
- the final product is stored at a controlled temperature of between 2-8°C, and the latest stability data showed that the drug product is stable under 4-8°C for 3 months and at accelerated condition of 25°C for 2 months and at 37°C for 2 weeks. Based on these test results at this time, the data supports a shelf life of 12 months and the stability program is on-going to support longer shelf life.
- the drug product is supplied in spray bottle with mechanical pump.
- the components of the container closure system are in conformance to the compendial requirements for spray bottle containers for pharmaceutical use.
- the drug product is produced in GMP qualified facilities under ISO5, ISO7, and ISO8 clean room classification.
- Stability was established according to ICH guideline for biological products Q1A and Q5C. Sponsor plans to test the drug product stability under 4-8°C for 36 months, at accelerated conditions 25°C for 6 months and 37°C for 1 month. Stability data for 3 representative batches of drug product 20200209, 20200210, and 20200311 is shown in the following Tables 14-17.
- Example 3 Treatment by rSIFN-co of COVID-19 patients with symptoms
- COVID-19 patients with symptoms such as mild pneumonia are treated by rSIFN-co or placebo for up to 10 to 28 days.
- rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time.
- Clinical improvement is determined by assessing radiological improvement and/or virus nucleic acid negative conversion at the end of treatment period. The median time to reach clinical improvement is approximately 8 to 16 days, and an overall rate of clinical improvement at the end of treatment is approximately 80%to 90%.
- Example 4 Treatment by rSIFN-co of COVID-19 patients without symptoms
- COVID-19 patients without symptoms are treated by rSIFN-co or placebo for up to 10 to 28 days.
- rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time. Any COVID-19 symptoms are assessed during the course of treatment.
- Patients treated with rSIFN-co are expected to show no symptom throughout the study, or significantly less treated patients show symptoms compared to patients receiving placebo.
- Example 5 Treatment by rSIFN-co in combination with other agents of COVID-19 patients with symptoms
- COVID-19 patients with symptoms such as mild pneumonia are treated by rSIFN-co in the presence of one or more agents for up to 10 to 28 days.
- rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time.
- one or more of the following agents are also administered: (1) lopinavir (twice a day, 200 mg each time) , (2) ritonavir (twice a day, 50 mg each time) , (3) arbidol or umifenovir (three times a day, 200 mg each time) , (4) Remdesivir (100 mg per day) , (5) hydroxychloroquine (200 mg per day) , and (6) dexamethasone (4 mg per day) .
- Clinical improvement is determined by assessing radiological improvement and/or virus nucleic acid negative conversion at the end of treatment period. The median time to reach clinical improvement and virus nucleic acid negative conversion is expected to be significantly shorter in patients receiving rSIFN-co than patients not receiving rSIFN-co.
- Example 6 Safety and tolerability in healthy subjects receiving rSIFN-co
- subjects receive rSIFN-co by nebulization or injection, or placebo for up to 10 to 28 days. Any symptoms or adverse events are assessed and recorded during the course of the study.
- Example 7 Treatment by rSIFN-co of COVID-19 patients on respirators
- COVID-19 patients on respirators are treated by rSIFN-co or placebo for up to 10 to 28 days.
- rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time.
- Clinical improvement is determined by assessing blood oxygen level during and at the end of treatment period. The median time to reach clinical improvement is approximately 8 to 16 days, and an overall rate of clinical improvement at the end of treatment is approximately 80%to 90%.
- Example 8 Treatment by rSIFN-co Spray
- healthy subjects or patients with an asymptomatic infection of Covid-19 receive 2 sprays per nostril and 4 sprays through pharynx once daily. Each spray comprises 2 ⁇ g of rSIFN-co.
- the period of treatment is approximately 5 days to 20 days.
- Subjects treated with rSIFN-co are expected to show no symptom throughout the study, or significantly less treated patients show symptoms compared to patients receiving placebo.
- each spray comprises 2 ⁇ g of rSIFN-co.
- the period of treatment is approximately 10 days to 30 days.
- the median time to reach clinical improvement and virus nucleic acid negative conversion is expected to be significantly shorter in patients receiving rSIFN-co than patients not receiving rSIFN-co.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Virology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Communicable Diseases (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Oncology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pulmonology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention discloses a method for preventing and/or treating COVID-19 (SARS-Cov-2) coronavirus infection by administering to a subject in need thereof a therapeutically effective dose of recombinant super-compound interferon (rSIFN-co) for a period. In one embodiment, the dose is from 1.0 to 100 μg per day and the period is at least 3 days and up to 30 days. In one embodiment, the rSIFN-co is administered by nasal spray, pharynx spray or intravenous injection.
Description
The present invention relates to recombinant super-compound interferon (rSIFN-co) and its use to treat COVID-19 patients with or without symptoms.
Since the outbreak of coronavirus disease 2019 (COVID-19) in December 2019, the disease has exploded into a global pandemic. As of December 8, 2020, nearly 66 million COVID-19 cases and 1,500,000 deaths have been reported [1] . From a report of 72, 314 COVID-19 patients in China, 81%of the cases were categorized into mild diseases (defined as no pneumonia or mild pneumonia) , 14%were severe (defined as dyspnea, respiratory frequency ≥30 breaths/min, SpO
2 ≤93%, PaO
2/FiO
2 <300 mmHg, and/or lung infiltrates >50%within 24 to 48 hours) , and 5%were critical (defined as respiratory failure, septic shock, and/or multiple organ dysfunction or failure) [2] . Common symptoms of COVID-19 include fever (83–99%) , cough (59–82%) , fatigue (44–70%) , anorexia (40–84%) , shortness of breath (31–40%) , and myalgias (11–35%) [3] . Other reported symptoms have included, but are not limited to, sore throat, nasal congestion, headache, diarrhea, nausea and vomiting, anosmia, and ageusia [3] . The mortality of COVID-19 varies by geographical area. According to a recent analysis conducted by the Centers for Disease Control and Prevention (CDC) of the United States, more than 1.3 million laboratory-confirmed cases were reported between January and May 2020. Of these cases, 14%of patients required hospitalization, 2%were admitted to an intensive care unit, and 5%died [4] . Although individuals of all ages are at risk of infection and severe disease, known risk factors for rapid deterioration, severe disease, and/or increased mortality include older age (more than 60 years) , noncommunicable diseases (e.g., diabetes, hypertension, cardiac disease, chronic lung disease, chronic kidney disease, immunosuppression and cancer) , and smoking [5] .
COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) , a newly discovered virus in the family of Coronaviridae (beta) , which has 89%nucleotide identity with bat SARS-like-CoVZXC21 and 82%with human SARS-CoV [6] . SARS-CoV-2 is an enveloped, positive-sense, single-stranded RNA virus. Virus entry is achieved through binding of viral spike (S) protein to its receptor, angiotensin-converting enzyme 2 (ACE2) , on the host cell [7] .
Current studies suggest that transmission of SARS-CoV-2 is primarily occurs through direct, indirect, or close contact with infected people via secretions such as respiratory droplets or saliva containing SARS-CoV-2 [3, 8] . According to the China National Health Commission’s guideline for diagnosis and treatment of COVID-19, virus could be found in human respiratory epithelial cells using in vitro culture system in about 96 hours [9] . Investigations of COVID-19 case clusters showed that transmission can occur from presymptomatic patients, from 1 to 3 days prior to symptom onset [10, 11] , which causes difficulties to contact tracing and disease control. As reports have suggested that viral shedding is highest at the time of symptom onset and that decreased viral load was correlated with reduced infectivity [12, 13] , transmissibility of COVID-19 may peak on or before symptom onset [14] . Therapeutics targeting virus reproduction may not only present with potential of disease treatment but also restricting its spreading.
To evaluate the efficacy of safety of nebulized rSIFN-co for moderate-to-severe COVID-19 patients, a phase II, randomized, single-blind study was conducted in China. Ninety-four (94) subjects were randomized in a 1: 1 ratio to receive either 12 million IU of rSIFN-co (twice daily) or 5 million IU of IFN alfa (twice daily) for a maximum of 28 days in addition to treatment with standard antiviral agents. It was found that subjects in the rSIFN-co arm had a significantly shorter time to clinical improvement (11.5 days vs 14.0 days) , radiological improvement (8.0 days vs 10.0 days) , and viral nucleic acid negative conversion (7.0 vs 10.0 days) than the IFN alfa arm. On Day 14, a significant higher proportion of patients receiving rSIFN-co had clinical improvement (65.2%vs 39.6%) . The percentage of subjects reporting adverse event during the study was lower in the rSIFN-co than the IFN alfa arm (28.3%vs 37.5%) [23] . However, there is no study suggesting whether rSIFN-co alone would treat COVID-19 infection, nor the effective dose of rSIFN-co for treating COVID-19 infection with no/limited side effects.
SUMMARY OF THE INVENTION
The present invention discloses a method that prevents and/or treats COVID-19 (SARS-Cov-2) coronavirus infection, including mild COVID-19 infection and asymptomatic infection, in a subject suffering from diseases in digestive system, liver, heart and/or kidney. The method comprises administering to the subject therapeutically effective doses, e.g., from 5 to 100 μg per day, of recombinant super-compound interferon (rSIFN-co) for a period of at least 3 days and up to 30 days. The administration can be by nasal spray or intravenous injection.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 is an illustration of the study schema.
Figure 2 compares structure of rSIFN-co and IFN-α2b
Figure 3 compares structure of rSIFN-co and Infergen of Amgen
Figure 4 shows vector diagram of rSIFN-co in Plasmid pHY-5.
Figure 5 shows a manufacturing flowchart for rSIFN-co drug product.
Currently, there are no specific treatment for COVID-19, and no known agent administered before or after exposure can prevent SARS-CoV-2 infection. Symptomatic and supportive treatment is still the mainstay for the disease. Given the highly contagious nature of COVID-19 and its devastating social and economic impacts, development of effective pharmacologic interventions for disease prevention and control is of urgent need.
With abilities of immunomodulation and interfering with viral replications, interferons (IFNs) have long been used as antiviral treatments. The preclinical studies of rSIFN-co have demonstrated its superiority of anti-SARS-CoV-2 capacity than conventional IFNs. Moreover, dysregulated IFNs was found in COVID-19 patients, implicating the role of IFN in COVID-19 pathogenesis [18] . Hence, the potential of rSIFN-co as a protective and therapeutic agent against SARS-CoV-2 infection is suggested.
Type I interferon for COVID-19
Impairment of type I interferon (IFN) response was discovered in severe and critical COVID-19 patients. No IFN-β and low IFN-α production/activity were observed and was associated with a persistent blood viral load and an exacerbated inflammatory response [17] . By blocking viral infection and eliciting immune responses, administration of IFNs at early stage of disease in patients or as protective agents in healthy subject may help prevent disease exacerbation or development [18] .
In the China National Health Commission’s guideline for diagnosis and treatment of COVID-19, 5 million IU or equivalence of IFN α is recommended for the treatment of COVID-19 at a frequency of two times a day, with a duration of no more than 10 days [9] . The application of IFN α/β for COVID-19 has also been suggested by others [19, 20] .
In a recent open-label study conducted in Hubei, China, 2415 subjects were treated with recombinant human IFN α nasal drops (2-3 drops/nostril/time; 4 times/day) for 28 days. No new-onset COVID-19 was observed in these subjects. While no flu-like symptoms was reported, some subjects experienced transient irritation of nasal mucosa (e.g., itching) , which was subsequently resolved without discontinuing study intervention [21] .
In another study, the efficacy and safety of IFN β-1a were evaluated in patients with severe COVID-19. In addition to the standard of care, the patients in the IFN group received subcutaneous injection of 12 million IU/mL of IFN β-1a three times per week for two weeks [22] . Although time to reach clinical response was not significantly improved, the proportions of subjects discharged on Day 14 were significantly higher in the IFN group [22] .
rSIFN-co for COVID-19
Recombinant super-compound interferon (rSIFN-co) is a product of patented technological research developed by Sichuan Huiyang Life Science and Technology Corporation. It is a recombinant form of the naturally occurring cytokine interferon-alpha (IFN-α) which has a modified spatial configuration. rSIFN-co has the same amino acid sequence as the Amgen product
(Interferon Alfacon-1) consisting of 167 amino acids (MCDLPQTHSLGNRRALILLAQMRRISPFSCLKDRHDFGFPQEEFDGNQFQKAQAISVLHEMIQQTFNLFSTKDSSAAWDESLLEKFYTELYQQLNDLEACVIQEVGVEETPLMNVDSILAVKKYFQRITLYLTEKKYSPCAWEVVRAEIMRSFSLSTNLQERLRRKE, SEQ ID NO: 1) . The amino acid homology of
and rSIFN-co are naturally occurring interferon α-2a and interferon α-2b, with only a difference of 18 and 19 amino acids, respectively. This amino acid difference results in rSIFN-co having 10 times greater affinity for IFN-specific cell surface receptors IFNAR1 compared to interferon α-2a and interferon α-2b [15] . rSIFN-co binds to IFN-specific cell surface receptors, resulting in the transcription and translation of genes whose protein products have antiviral, antiproliferative, anticancer, and immune-modulating effects. The 3-dimensional conformational change improves efficacy and causes fewer side effects compared to IFN-a. The nucleic acid sequence encoding the amino acid sequence of SEQ ID NO. 1 is as follows (SEQ ID NO: 2) .
In one embodiment, the rSIFN-co can be produced by the method comprising the following steps: introducing a nucleotide sequence comprising SEQ ID NO: 2 that encodes the recombinant interferon into an isolated host cell; culturing the host cell under appropriate condition for expression of the recombinant interferon; and harvesting the recombinant interferon, wherein the recombinant interferon has an amino acid sequence of SEQ ID NO: 1, and the recombinant interferon inhibits secretion of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) of Hepatitis B Virus. Further, the host cell is Escherichia coli. Further, the nucleotide sequence comprising SEQ ID NO: 2 is under the control of the promoter PBAD. Further, the harvesting step comprises extraction of the interferon from the fermentation broth, collection of the inclusion bodies, denaturation and renaturation of the harvested interferon. Still further, the harvesting step also comprises separation and purification of the recombinant interferon. Met at the N-terminus of SEQ ID NO: 1 is resected in mature protein.
In one embodiment, this invention provides a crystalline recombinant interferon comprising the amino acid sequence of SEQ ID NO: 1. Further, this crystal belongs to the trigonal system. In one embodiment, the space group of this crystal is P3
121. In some embodiments, the unit cell parameters of this crystal are
α = β = 90°, γ = 120°, with a variability of at most 5%in all cell parameters. In some embodiments, said crystal contains two molecules in one asymmetric unit. In some embodiments, said crystal comprises covalently or non-covalently bound metal ions. Further, said metal ions can be magnesium ion, zinc ion and the like, these metal ions can mediate the formation of the interferon dimers in the crystal.
In an in vitro study, the anti-SARS-CoV-2 activities of rSIFN-co has been investigated in Vero-E6 cell to assess rSIFN-co’s inhibitory effect on 2019-nCoV’s replication at cell level. The Vero-E6 cell line was kept in the lab at a multiplicity of infection (MOI) of 0.005. The study used reagents including DMEM (Gibco) , fetal calf serum (Gibco) , DMSO (sigma) , double-antibody, pancreatine, etc., kits including Cell Counting Kit-8 (CCK-8) (B34304, bimake) , QIAamp viral RNA mini kit (52906, Qiagen) , Novel Coronavirus (2019-nCoV) Real Time RT-PCR kit (by Wuhan Institute of Virology) , and main instruments such as Varioskan Flash (by Thermo Fisher Scientific) , QIAcube HT 9001793 (by Qiagen) and CFX96 Touch Real-Time PCR Detection System (by Bio-rad) .
The following method was used to determine the cytotoxicity of rSIFN-co and recombinant human Interferon α-2b injection (pseudomonas) :
a) Grow Vero E6 cells in 96 micro well plate one day in advance, 1×10
4 cells per well (Attention: The peripheral wells on the plate shall not be used for experiment, and add in PBS to prevent the volatilization of medium in other wells; )
b) Observe the cells and when the confluence reaches 50%, use 2×DMEN with 4%FBS to dilute rSIFN-co 2.17 times to obtain the maximum concentration, i.e., 6×10
8 pg/ml, and dilute recombinant human Interferon α-2b injection (pseudomonas) 1.5 times to obtain the maximum concentration, i.e., 2×10
7 pg/ml. Then use DMEN with 2%FBS for 2 times gradient dilution of rSIFN-co, recombinant human Interferon α2b injection (pseudomonas) and Remdesivir respectively. The final set of concentrations of rSIFN-co was 6×10
8, 3×10
8, 1.5×10
8, 7.5×10
7, 3.75×10
7, 1.875×10
7 and 0 pg/ml, the final set of concentrations of recombinant human Interferon α-2b injection was 2×10
7, 1×10
7, 5×10
6, 2.5×10
6, 1.25×10
6, 6.25×10
5 and 0 pg/ml, and the final set of concentrations of Remdesivir was 320, 160, 80, 40, 20, 10 and 0 μM;
c) Add the above solutions to the plate at 100 μL/well, and 4 wells per concentration. Set control group (drug free) and blank group (cell free) . Put plates into an incubator with 5%CO
2 at 37 ℃; and
d) After adding drug 48 hours, add Cell Counting Kit-8 (CCK-8) at ratio of 1/10 cell culture medium and mix well, avoiding bubbles. Put plates into the incubator at 37 ℃ for about one hour until it turns orange. Zero setting microplate reader with blank group, then read OD
450 values of the whole plate. Calculate the survival rate by the formula: Survival rate = Drug group OD
450 /Control group OD
450 × 100%, and then calculate the half cytotoxicity concentration (CC
50) of drugs.
Results are summarized in Table 1.
TABLE 1. The Half Cytotoxicity Concentration (CC
50) of rSIFN-co and Recombinant Human Interferon α2b Injection (Pseudomonas) to Vero E6 Cells
The following method was used to evaluate the inhibitory effect of rSIFN-co and recombinant human Interferon α-2b injection (pseudomonas) on 2019-n-CoV:
a) Grow Vero-E6 cells in a 24-well plate, 8×10
4 cells per well. Put the plate into 37 ℃incubator with 5%CO
2. When the confluence reaches 70%-80%, using DMEN with 2%FBS for 2 times gradient dilution of rSIFN-co, recombinant human Interferon α-2b injection (pseudomonas) and Remdesivir, respectively. Discard the culture medium in each well and add in 1 ml 2019-nCoV (COVID-19) solution (MOI as 0.005) and the DMEM with drugs (set the concentrations of Remdesivir as 10, 5, 2.5, 1.25, 0.625, 0.3 and 0 μM, concentration of rSIFN-co as 60, 30, 15, 7.5, 3.75 and 0 pg/ml, and concentration of recombinant human Interferon α-2b injection (pseudomonas) as 300, 150, 75, 37.5, 18.75 and 0 pg/ml) . Set a control group at the same time. Collect supernatant 24 hours after infection;
b) Use real time RT-PCR (qRT-PCR) for RNA quantitative detection on collected virus. Take 20 μL collected supernatant after infection and extract RNA following instructions of QIAamp viral RNA mini kit. Conduct the qRT-PCR by using Novel Coronavirus (2019-nCoV) Real Time RT-PCR kit (TaqMan probe method) ;
c) Calculate the drug inhibition rate at each concentration. Inhibition rate (%) = 1 -Viral RNA Copies in test groups /Viral RNA Copies in drug free group × 100%. Analyze the half effective concentration of drugs (EC
50) by GraphPad PrisM6.0; and
d) Calculate the therapeutic index of drugs by formula: Therapeutic Index (TI) = half cytotoxicity concentration (CC
50) /half effective concentration (EC
50) .
Results are summarized in Tables 2 and 3.
Table 2. Half Effective Concentration (EC
50) of rSIFN-co and Recombinant Human Interferon α2b Injection (Pseudomonas) for Inhibiting 2019-nCoV (COVID-19) Replication in Vero-E6 Cell Model
Table 3. Therapeutic Index (TI) of rSIFN-co and Recombinant Human Interferon α2b Injection (Pseudomonas)
In summary, the study showed that the EC
50 and CC
50 of rSIFN-co were 14.6 pg/mL and 7.12×10
8 pg/mL, respectively, whereas the EC
50 and CC
50 of IFN α-2b were 162 pg/mL and 1.37×10
7 pg/mL, respectively. The therapeutic index (TI=CC
50/EC
50) was 4.88×10
7 for rSIFN-co and 8.46×10
4 for IFN α-2b. The results suggested that rSIFN-co exhibited a higher efficacy against SARS-CoV-2 and a lower cytotoxicity as compared with IFN α-2b. In a separate study, rSIFN-co further exhibited a lower EC
50 at a MOI of 0.05 and 0.5 than IFN α-2b or IFN β-1b (EC
50 when the MOI=0.05 for rSIFN-co: 133.75 pg/mL, IFN α-2b: 575.65 pg/mL, IFN β-1b: 194.60 pg/mL; EC
50 when the MOI=0.5 for rSIFN-co: 164.15 pg/mL, IFN α-2b: 2432.50 pg/mL, IFN β-1b: 1240.00 pg/mL) .
In a pharmacokinetic (PK) study comparing rSIFN-co and
21 healthy male volunteers were enrolled at the Drug Clinical Research Base, West China Hospital of Sichuan University. The subjects were given a single dose of 9 μg rSIFN-co (Group A) or 9 μg
(Group B) , and later switched over, and their blood taken at specified intervals. The PK results suggest that rSIFN-co induces more 2′-5′-oligoadenylate synthetase (2′, 5′-OAS; a specific marker triggered by interferon) over a longer time period, when compared to 2′, 5′-OAS induced by
In addition, the incidence of adverse events in the rSIFN-co group was lower than that in the
group. The majority of the adverse effects experienced after the treatment of both rSIFN-co and
included fever, headache, myalgia, fatigue and chills.
Type I interferons are known to play an important role in first-line defense against viruses by stimulating the expression of proteins such as ribonucleic acid (RNA) -dependent protein kinase, 2′, 5′-OAS, RNase L, and RNA-specific adenosine deaminase [16] . In the phase II clinical study of rSIFN-co in patients with chronic hepatitis B, rSIFN-co at 9 μg exhibited similar efficacy as 50 μg interferon α-1b, which was demonstrated by the suppression of hepatitis B virus replication, HBeAg/HBeAb seroconversion, and ALT reversion to normal levels. The rSIFN-co treatment was also better tolerated compared to interferon α-1b.
A set of experiments was also conducted to compare rSIFN-co, IFN α2b, β-1b, and hydrochloroquine at a high Multiplicity of Infection (MOI) of 0.5. The TI of rSIFN-co was is 4.34×10
6 which is 770 times higher than that of IFN α2b; EC50 for rSIFN-co is 164.15 pg/mL which is 7 times lower in concentration as compare to β-1b which requires 1240.00 pg/mL. The result is summarized in Table 4.
Table 4. Comparison of rSIFN-co, INFα2b, IFN β-1b, and Hydrochloroquine in MOI of 0.5
An in vitro study was conducted to compare rSIFN-co protection in viral infection to various forms of commercially available IFN products and Remdesivir. This includes IFN-α2b, IFNα8, IFNα14, IFNβ-1b, IFNω. As shown in Table 5, this study demonstrated that rSIFN-co at a concentration of 50U/mL is effective in protecting viral infection of up to 48 hours. This study further demonstrated that rSIFN-co could induce 2’-5’-oligoadenylate synthetase (2’, 5’-OAS) and under the same dose could have a better protective mechanism.
Table 5. Infection Protection comparing rSIFN-co to various commercial IFNs
Furthermore, during the outbreak of severe acute respiratory syndrome (SARS) , rSIFN-co was distributed with the consent of the Sichuan (aprovince in China) working group on SARS prevention and control. rSIFN-co spray was allocated to 3,000 users, including doctors and nurses in hospitals, populated areas with a high risk for SARS, and the National research group. Among all users, the highest dose was 16 million international unit (IU) and the longest treatment duration was over two months. No side effects connected to the use of the spray was reported, and none of users administered with rSIFN-co spray has been infected by SARS.
In one embodiment, the present invention provides a method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject. In one embodiment, the method comprises administering to the subject a therapeutically effective dose of recombinant super-compound interferon (rSIFN-co) for a period of at least 3 days.
In one embodiment, the rSIFN-co is administered to said subject by nasal spray, pharynx spray or intravenous injection.
In one embodiment, the therapeutically effective dose ranges from 5 μg to 100 μg per day.
In one embodiment, the therapeutically effective dose is selected from the group consisting of 8 μg, 16 μg, 32 μg, 64 μg and 96 μg per day.
In one embodiment, the period is 5 to 30 days.
In one embodiment, the subject suffers from one or more diseases in one or more of the digestive system, liver, heart, and kidney.
In one embodiment, the subject receives one or more additional treatments selected from the group consisting of Remdesivir, hydroxychloroquine, and dexamethasone.
In one embodiment, the rSIFN-co forms a crystal of the space group of P3
121, and the unit cell parameters of the crystal are
α=β=90°, and γ=120°.
In one embodiment, the subject is healthy or has an asymptomatic infection of Covid-19 coronavirus, and said subject receives 2 sprays per nostril and 4 sprays through pharynx once daily, each spray comprising 0.2-5.0 μg of said rSIFN-co.
In one embodiment, the subject shows mild covid-19 infection and receives 2 sprays per nostril and 4 sprays through pharynx twice daily, each spray comprising 0.2-5.0 μg of said rSIFN-co.
In one embodiment, the rSIFN-co has a specific activity greater than 5.0 x 10
8 IU/mg protein.
In one embodiment, the nasal spray, pharynx spray or intravenous injection comprises one or more of stabilizer, bacteriostat, surfactant, metal complex, isosmotic adjusting agent, buffering agent, and viscosity modifier.
In one embodiment, the stabilizer is human serum albumin. In one embodiment, the metal complex is disodium edetate. In one embodiment, the surfactant is Tween 80. In one embodiment, the buffering agent is citric acid. In one embodiment, the viscosity modifier is glycerol. In one embodiment, the isosmotic adjusting agent is sodium chloride. In one embodiment, the bacteriostat is benzyl alcohol.
In one embodiment, the nasal spray, pharynx spray or intravenous injection has a pH value of about 5.0.
In one embodiment, the nasal spray or pharynx spray is stable under 4-8℃ for 3 months, at 25℃ for 2 months and at 37℃ for 2 weeks.
In one embodiment, the present provides a method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject. In one embodiment, the method comprises administering to the subject a composition comprising a therapeutically effective dose of recombinant super-compound interferon (rSIFN-co) for a period of at least 3 days
In one embodiment, the present provides a method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject by administering a composition consisting of a therapeutically effective dose of rSIFN-co for a period of at least 3 days.
Objectives
The primary objective is to assess the safety and tolerability of rSIFN-co in healthy subjects in close contact with confirmed COVID-19 case (s) and patients with mild COVID-19 or asymptomatic infection.
The secondary objective is to assess the efficacy of rSIFN-co in healthy subjects in close contact with confirmed COVID-19 case (s) and patients with mild COVID-19 or asymptomatic infection.
The primary endpoint is the safety and tolerability profiles of rSIFN-co.
The secondary endpoint for Healthy subjects in close contact with confirmed COVID-19 case (s) is:
1. Incidence of laboratory-confirmed SARS-CoV-2 infection over 28 days (Day 1 to Day 28)
2. Severity and clinical status of COVID-19 over the study period, and
3. Incidence of COVID-19-related complications.
The secondary endpoint for patients with mild COVID-19 or asymptomatic infection is:
1. Proportion of subjects with COVID-19-related hospitalization by Day 28,
2. Proportion of subjects who require supplemental oxygen or mechanical ventilation by Day 28,
3. Clinical improvement rates on Days 5 and 10,
4. Change from baseline per FDA definition ordinal scale on Days 5 and 10,
5. Time to clinical improvement from the lowest outcome scores,
6. Change in disease severity from baseline to Days 5 and 10,
7. Qualitative and quantitative polymerase chain reaction (PCR) for SARS-CoV-2 in respiratory tract on Days 1, 2, 3, 5, 7, 10, 15, and 21,
8. Time to clearance of SARS-CoV-2, defined as 2 consecutive negative swabs (sampling interval ≥ 24 hours) , and
9. Time to resolution of all symptoms present at baseline.
The exploratory endpoint for Healthy subjects in close contact with confirmed COVID-19 case (s) is the subjects’ evaluation on ease of use, discomfort using the study product, and overall satisfaction.
The exploratory endpoint for patients with mild COVID-19 or asymptomatic infection is:
1. Incidence of COVID-19 pneumonia, cough, and fever over the study period,
2. Proportion of subjects without fever on Days 1, 5, 10, and 15,
3. Subjects’ evaluation on ease of use, discomfort using the study product, and overall satisfaction,
4. Incidence of admission to a critical care unit, and
5. COVID-19 seroconversion rate for IgG and IgM on Days 5, 10, 15, and 21.
STUDY DESIGNS
General Design
This is a phase II, multi-site, randomized, double-blind, placebo-controlled pilot study that aims to evaluate the safety and preliminary efficacy of rSIFN-co in healthy subjects in close contact with confirmed COVID-19 case (s) as well as patients with mild COVID-19 or asymptomatic infection. Approximately 120 subjects are planned to be enrolled, including 60 healthy subjects and 60 patients.
For the healthy subjects in close contact with confirmed COVID-19 case (s) (Group A) , approximately 60 subjects will be randomized in a 1: 1 ratio to one of the two arms: placebo arm or rSIFN-co arm.
For the patients with mild COVID-19 or asymptomatic infection (Group B) , approximately 60 subjects will be randomly assigned in a 1: 1 ratio to receive standard of care (SoC) plus placebo or SoC plus rSIFN-co.
All eligible subjects will receive the study medications for 10 days, around 8 million IU (16 μg) of rSIFN-co for healthy subjects and 16 million IU (32 μg) for COVID-19 patients daily (see Dosage, Preparation and Administration of Investigational Product for details) . After 10 days of treatment, a safety follow-up visit on Days 15, 21, 28, and 60 will be conducted. The safety and efficacy parameters will be monitored throughout the study period.
Scientific Rationale for Study Design
To evaluate the safety and preliminary efficacy of the investigational product in healthy subjects in close contact with confirmed COVID-19 case (s) and patients with mild COVID-19 or asymptomatic infection, the study is designed as a randomized, double-blind, placebo-controlled study to minimize the potential for subjective bias. The inclusion of placebo arms will provide a control for comparison of efficacy endpoints between the treatment groups. The study will be conducted at multiple study centers to facilitate subject enrollment and to increase the generalizability of the study results.
As therapeutic options for patients with mild COVID-19 are limited, and symptomatic and supportive treatment is currently the mainstay for COVID-19. For this reason, local SoC for COVID-19 is incorporated and permitted for patients with mild COVID-19 or asymptomatic infection (Group B) in order to satisfy subjects’ need.
Justification for Dose
The doses evaluated for this study, 8 and 16 million IU, are based on the totality of the data from previous trials in adults.
In the previous phase I study conducted at the National Cancer Centre in Singapore, there were no dose-limiting toxicity observed in subjects subcutaneously administered with rSIFN-co at doses of up to 30 μg (refer to the IB for details) . Additionally, the phase II study evaluating the efficacy and safety of rSIFN-co in moderate-to-severe COVID-19 patients also demonstrated that nebulized rSIFN-co (12 million IU, twice daily) was safe and well tolerated [23] . With a dose of 12 million IU twice per day, no SAEs were reported and all AEs were classified as either grade 1 or 2 in patients treated with rSIFN-co. As the present study include only healthy subjects or mild/asymptomatic COVID-19 patients, the daily dose of rSIFN-co is reduced to 8 and 16 million IU, respectively. The selected doses are considered adequate with its known safety profile.
Inclusion Criteria
An eligible subject must fulfill
ALL of the following inclusion criteria:
1. Male or females ≥ 18 years of age at the time of informed consent;
2. Willing and able to provide written informed consent for the trial;
3. Healthy subjects in close contact with confirmed COVID-19 case (s) or patients with mild COVID-19 or asymptomatic infection;
a. Healthy subjects in close contact with confirmed COVID-19 case (s) ;
I. With exposure to confirmed COVID-19 case (s) within 96 hours (refer to the definition of healthy subjects in close contact with confirmed COVID-19 case [s] ) ,
II. Negative reverse transcriptase polymerase chain reaction (RT-PCR) test for SARS-CoV-2 infection, with the sample collected within 72 hours prior to screening,
III. Have no previous confirmed COVID-19 diagnosis,
IV. Without symptoms of respiratory infection, including fever, cough, fatigue, anorexia, shortness of breath, myalgias, sore throat, nasal congestion, headache, gastrointestinal symptoms, loss of smell (anosmia) , loss of taste (ageusia) , or other symptoms associated with COVID-19, AND
V. With stable health status, as judged by the investigator and determined by physical examination, vital signs, medical history, and laboratory tests at screening;
b. Patients with mild COVID-19 or asymptomatic infection;
I. Meeting the criteria of mild COVID-19 or asymptomatic infection (refer to the definition of COVID-19 disease severity) ,
II. Positive RT-PCR test for SARS-CoV-2 infection, with the sample collected within 72 hours prior to screening, AND
III. Without evidence of viral pneumonia or hypoxia;
4. Women of childbearing potential must have a negative pregnancy test result at screening; and
5. Males and females who are fertile must adhere to contraception requirements for the duration of the study.
Definitions
The following terms, including abbreviations listed in Table 6 below, shall be used to describe the present invention. In the absence of a specific definition set forth herein, the terms used to describe the present invention shall be given their common meanings as understood by those of ordinary skill in the art.
TABLE 6. LIST OF ABBREVIATIONS
Healthy subjects in close contact with confirmed COVID-19 case (s) : Subjects with exposure to confirmed COVID-19 case (s) within 96 hours and belong to one of the following exposure situations per health department directives:
1. Staying together in the same room with confirmed COVID-19 case (s) , e.g., living, studying, working, dining and hanging out with confirmed COVID-19 case (s) , etc.
2. Taking the same transport with confirmed COVID-19 case (s) , e.g., all personnel in the cabin of civil aircraft; all passengers in the same carriage of a fully enclosed air-conditioned train and the crew members serving for this area; all personnel of fully sealed air-conditioned vehicles; all personnel in the same cabin of the ship and crew members serving for that cabin.
3. Medical staffs below without any protection of level 2 or above.
(1) People mainly working in the emergency room (e.g., doctors, nurses, assistants, triage personnel) ;
(2) People mainly working in intensive care unit (ICU) (e.g., physicians, nurses, support staff, respiratory therapists) ;
(3) People performing aerosol generation procedures (i.e., anesthesiologist, cRNA) ;
(4) First responders (i.e., first aid personnel, paramedics) ; and
(5) People working in the Department of general medicine, pulmonary medicine, infectious diseases, and isolation wards.
Note: Level 2 protection refers to wearing a disposable cap, medical protective mask (N95 or higher level medical protective mask) , goggles (anti-fog type) or protective mask (anti-fog type) , medical protective suit or overalls (white gown) , disposable gloves and disposable shoe covers.
COVID-19 disease severity (CDC definitions) [25]
- Asymptomatic or presymptomatic infection: Individuals who test positive for SARS-CoV-2 by virologic testing using a molecular diagnostic (e.g., polymerase chain reaction) or antigen test, but have no symptoms.
- Mild disease: Individuals who have any of the various signs and symptoms of COVID-19 (e.g., fever, cough, sore throat, malaise, headache, muscle pain) and a SpO
2 ≥94%on romm air at sea level, without shortness of breath, dyspnea, or abnormal chest imaging.
- Moderate disease: Individuals who have evidence of lower respiratory disease by clinical assessment or imaging and SpO
2 ≥94%on room air at sea level.
- Severe disease: Individuals who have respiratory frequency >30 breaths per minute, SpO
2 <94%on room air at sea level, ratio of arterial partial pressure of oxygen to fraction of inspired oxygen (PaO
2/FiO
2) <300 mmHg, or lung infiltrates >50%.
- Critical disease: Individuals who have respiratory failure, septic shock, and/or multiple organ dysfunction.
Dosage, Preparation and Administration of Investigational Product
The subjects will receive rSIFN-co or placebo, for a maximum of 10 to 28 days. For combination therapy, rSIFN-co will be administered with one or more other agents, wherein the control group will receive said one or more other agents only.
The dose of rSIFN-co given by nebulization or injection is equivalent to 8 to 16 million IU (32 μg) per day. Other agents will be given in their usual dosages.
Treatment Period
Healthy subjects (without symptoms) in close contact with confirmed COVID-19 case (s)
● Perform physical examination;
● Assess vital signs (including SpO
2) ;
● Document COVID-19-related symptoms, complications, clinical status, and disease severity, if applicable;
● Randomize eligible subjects into one of the two treatment arms (i.e., placebo or rSIFN-co) ;
● Collect respiratory tract sample (s) for viral RNA quantification and qualification;
● Dispense study medications and diary card; and
● Record AEs and concomitant medications;
● Review how to complete diary with subject.
● Perform physical examination;
● Assess vital signs (including SpO
2) ;
● Document COVID-19-related symptoms, complications, clinical status, and disease severity, if applicable;
● Perform ECG if clinically indicated;
● Collect blood samples for laboratory tests;
● Collect respiratory tract sample (s) for viral RNA quantification and qualification;
● Collect blood samples for serologic test;
● Review electronic diary entries with subject prior to leaving clinic, including any AE/SAE information and have subject correct any discrepancies; and
● Record AEs and concomitant medications.
Day 10
● Perform physical examination;
● Assess vital signs (including SpO
2) ;
● Document COVID-19-related symptoms, complications, clinical status, and disease severity, if applicable;
● Obtain chest imaging;
● Perform ECG if clinically indicated;
● Collect blood samples for laboratory tests;
● Collect blood, stool, and respiratory tract sample (s) for viral RNA quantification and qualification;
● Collect blood samples for serologic test;
● Review electronic diary entries with subject prior to leaving clinic, including any AE/SAE information and have subject correct any discrepancies;
● Complete questionnaire for ease of use and discomfort using the study product; and
● Record AEs and concomitant medications.
COVID-19 patients with mild symptoms or asymptomatic infection
● Perform physical examination;
● Assess vital signs (including SpO
2) ;
● Document COVID-19-related symptoms, complications, clinical status, and disease severity;
● Randomize eligible subjects into one of the two treatment arms (i.e., SoC+placebo or SoC+rSIFN-co) ;
● Collect respiratory tract sample (s) for viral RNA quantification and qualification;
● Dispense study medications and diary card; and
● Record AEs and concomitant medications.
● Perform physical examination;
● Assess vital signs (including SpO
2) ;
● Document COVID-19-related symptoms, complications, clinical status, and disease severity;
● Perform ECG if clinically indicated;
● Collect respiratory tract sample (s) for viral RNA quantification and qualification; and
● Record AEs and concomitant medications.
● Perform physical examination;
● Assess vital signs (including SpO
2) ;
● Document COVID-19-related symptoms, complications, clinical status, and disease severity;
● Obtain chest imaging;
● Perform ECG if clinically indicated;
● Collect blood samples for laboratory tests;
● Collect respiratory tract sample (s) for viral RNA quantification and qualification;
● Collect blood and stool samples for viral RNA quantification and qualification (only for Day 10) ;
● Collect blood samples for serologic test;
● Collect diary card and used/unused study medications (Day 10) ;
● Review electronic diary entries with subject prior to leaving clinic, including any AE/SAE information and have subject correct any discrepancies;
● Record AEs and concomitant medications; and
● Complete questionnaire for ease of use and discomfort using the study product (only for Day 10) .
Follow-up Visits (Days 15, 21, 28 and 60)
● Perform physical examination;
● Perform pregnancy test Day15;
● Assess vital signs (including SpO
2) ;
● Document COVID-19-related symptoms, complications, clinical status, and disease severity, if applicable;
● Obtain chest imaging, if clinically indicated;
● Perform ECG, if clinically indicated;
● Collect blood samples for laboratory tests on Day 15, thereafter only if clinically indicated;
● Collect respiratory tract sample (s) for viral RNA quantification and qualification;
● Collect blood and stool samples for viral RNA quantification and qualification (only for Day 28) ;
● Collect blood samples for serologic test (for Group A, the samples will only be collected on Day 15 and/or Day 28, if a positive RT-PCR result for SARS-CoV-2 is obtained in the preceding test) ;
● Review electronic diary entries with subject prior to leaving clinic, including any AE/SAE information and have subject correct any discrepancies;
● Collect electronic diary on Day 60
● Record AEs and concomitant medications.
After Day 60
Subjects with positive RT-PCR results on Day 60 will be continuously followed every two (2) weeks until a negative result is obtained.
Vital Signs
Vital sign measurements include blood pressures (systolic blood pressure [SBP] and diastolic blood pressure [DBP] ) , pulse rate, respiratory rate, body temperature, and SpO
2. Vital signs will be assessed at each visit. Results will be documented in the CRF. Measurements of all vital signs shall be completed in the during the visit within a time frame of 2 hours. Measurements should be taken in a seated position after resting for approximately 10 minutes. The same thermometer should be used consistently throughout the study. Subjects in Group B will be provided with an oximeter to help monitor their oxygen level. The subjects will be instructed to document daily oxygen level on the diary card.
Fever is defined as rectal, ear, or forehead temperature greater than 38 ℃. Site personnel should measure ear temperature of subject if possible. If other methods are used, the same method should be used consistently throughout the study.
Physical Examinations
A complete physical examination will be conducted by the Investigator or designee at screening. Complete physical examination items include general appearance, HEENT (head, eyes, ears, nose, and throat) , mouth, skin, neck (including thyroid) , lymph nodes, spine, cardiovascular system, respiratory system, gastro-intestinal system, nervous system, musculoskeletal system, blood and blood forming organs, mental status, and other body systems if applicable for describing the status of subject’s health. At subsequent visits, an abbreviated physical examination comprising a brief medical history and targeted physical examination will be performed at the discretion of the Investigator to evaluate possible adverse events.
The physical examination performed at screening will serve as the baseline. Any significant physical examination findings after dosing will be regarded as adverse events.
Electrocardiogram (ECG)
Single 12-lead ECG will be obtained at screening using an ECG machine that automatically calculates the heart rate and measures PR, QRS, QT, and QTc (either Bazette or Fridericia formulas) intervals. The assessment will follow the current SOPs at each participating study center. Throughout the study, additional ECGs may be obtained if clinically indicated. The Investigators will compare ECGs to the screening ECG and document overall interpretation of specific ECGs as either normal, abnormal with clinical significance, or abnormal without clinical significance. An AE will be recorded if clinically significant.
Laboratory Evaluations
Laboratory tests (including hematology, biochemistry, and coagulation) will be performed at screening, Days 5, 10, 15, 21, 28, and 60. All samples will be analyzed by the study center’s local laboratory per the standard operating procedures (SOPs) of the laboratory. The examination items will include analytes in the following table:
Additional tests may be conducted at any time during the study as determined necessary by the Investigator. A clinically significant change from baseline will be recorded as an AE.
SARS-CoV-2 Viral Load
Blood, stool and respiratory tract samples will be collected at scheduled visits. For respiratory tract samples, nasopharyngeal swabs are preferred [26] , but if these are not obtainable or additional samples are required by local regulations, oropharyngeal or nasal swabs may be obtained if necessary.
Samples will be delivered to the laboratories selected by the Sponsor to quantitatively and qualitatively detect viral RNA via RT-PCR assays approved by local regulatory authorities. Preparation, handling, storage, shipment, and examinations of specimens will follow the lab SOPs.
SARS-CoV-2 Serologic Test
For detection of immunoglobulin (Ig) M and IgG to SARS-CoV-2, blood samples will be collected at screening and on Days 5, 10, 15, and 28. Validated serologic tests for SARS-CoV-2 that are approved by local regulatory authorities will be applied and will be performed by the laboratories selected by the Sponsor. Preparation, handling, storage, shipment, and examinations of specimens will follow the lab SOPs.
Chest Imaging
Thoracic CT-scan or chest X-ray will be performed at Screening, Day 5 (only for mild COVID-19 patients or asymptomatic infection) , and End of Treatment (Day 10) . A standardized framework will be used to assess the chest imaging at each study site.
WHO Ordinal Scale
COVID-19-related clinical status will be assessed at each study visits. The WHO ordinal scale as enumerated in the below table will be applied to evaluate improvement in clinical status [27] .
*If hospitalized for isolation only, record status as for ambulatory patient.
Clinical improvement will be defined as a decrease of at least one point on the 11-point scale compared to the baseline value (e.g., from 2 to 1; from 2 to 0) .
COVID-19 Disease Severity and Symptoms
COVID-19 disease severity and symptoms will be assessed and documented at screening and throughout the entire study. Typical symptoms for patients with mild disease include fever, cough, fatigue, anorexia, shortness of breath, myalgias, sore throat, nasal congestion, headache, gastrointestinal symptoms, loss of smell (anosmia) , loss of taste (ageusia) , and without evidence of viral pneumonia or hypoxia.
The following COVID-19 disease severity will be applied:
● Asymptomatic or presymptomatic infection: Individuals who test positive for SARS-CoV-2 by virologic testing using a molecular diagnostic (e.g., polymerase chain reaction) or antigen test, but have no symptoms;
● Mild disease: Individuals who have any of the various signs and symptoms of COVID-19 (e.g., fever, cough, sore throat, malaise, headache, muscle pain) and a SpO2 ≥94%on room air at sea level, without shortness of breath, dyspnea, or abnormal chest imaging;
● Moderate disease: Individuals who have evidence of lower respiratory disease by clinical assessment or imaging and SpO2 ≥94%on room air at sea level;
● Severe disease: Individuals who have respiratory frequency >30 breaths per minute, SpO2 <94%on room air at sea level, ratio of arterial partial pressure of oxygen to fraction of inspired oxygen (PaO
2/FiO
2) <300 mmHg, or lung infiltrates >50%; and
● Critical disease: Individuals who have respiratory failure, septic shock, and/or multiple organ dysfunction.
The present invention will be better understood by reference to the examples as follows, but those skilled in the art will readily appreciate that the specific examples detailed are only for illustrative purpose, and are not meant to limit the present invention as described herein, which is defined by the claims following thereafter.
Throughout this application, various references or publications are cited. Disclosures of these references or publications in their entireties are hereby incorporated by reference into the application in order to more fully describe the state of the art to which this invention pertains. It is to be noted that the transitional term “comprising” , which is synonymous with “including” , “containing” or “characterized by” , is inclusive or open-ended, and does not exclude additional, un-recited elements or method steps.
EXAMPLES
Example 1: Preparation of rSIFN-co
The Master Cell Bank (MCB) and Working Cell Bank (WCB) for rSIFN-co are manufactured under cGMP. The DS manufacture process involves bioproduction and harvest then purification of rSIFN-co from the E. coli inclusion bodies using upstream and downstream manufacturing processing typical for this type of production.
To create rSIFN-co, the rSIFN-co cDNA was cloned into the E. coli high expression vector pHY-4 plasmid to yield the final recombinant plasmid pHY-5 (Figure 4) . Typical production and purification can be found in US Patent No. 8,415,151 and Patent No. 8,846,025.
Vector components in Figure 4 are:
● PBAD promoter, which derived from bacterial arabinose operon (araBAD operon) and was mainly regulated by CAP-cAMP and L-arabinose binding protein AraC i.e. product of ara Cgene
● Regulatory sequences araI, araO1 and araO2 of araBAD operon.
● araC gene coded PBAD promoter modulin –AraC.
● Ribosome binding site in the downstream of transcription start site, where PBAD promoter acted, followed by NdeI restriction enzyme cutting site.
● Transcription termination signal (rrnBT1T2)
● β-Lactamase coded by Bla gene could provide ampicillin resistance.
● ColE1 replicon determined the copy number and replication of plasmid in bacteria.
Fifty vials of MCB were manufactured and tested to ensure clone purity, stability, absence of adventitious agents and stable protein expression. The WCB was tested for mycoplasma, viruses, and non-host cell contamination. Additional tests were performed to confirm rSIFN-co identity via peptide map analysis and testing of the protein potency.
Animal-based ingredients, tryptone and acid hydrolyzed casein, are used in upstream manufacture and are of GMP quality and certified that they are sourced in a controlled and validated way as to reduce the risk of contamination or exposure to any diseases, especially Bovine Spongiform Encephalopathy.
The release testing specifications for the DS include protein content, biological activity, specific activity, purity, molecular weight, exogenous DNA, host protein, antibiotic residue, endotoxin and a number of characterization assays.
The identity of the primary sequence of rSIFN-co was confirmed using MS analysis. Mass-Spectrometry analysis was performed using MALDI-TOF-MS, and the results were consistent with the theoretical value of rSIFN-co minus methionine (19434.28Da) and reported in references (19434Da) . Traditional N-terminal sequencing by Edman degradation confirmed the identity of the first 15 amino acids. The identity was also confirmed by isoelectric focusing. In this gel method, the overall charge state of rSIFN-co (pI) was determined which is partly a measure of the number of charged amino acids in the molecule (primary structure) and partly the influence of structural considerations (secondary structure) around those charged residues. Secondary structure analysis was accomplished by CD Spectroscopy. This analysis also showed identical thermal structural stability.
The biological activity of rSIFN-co was determined based on the effect of rSIFN-co to protect cells against a viral cytopathic effect. It is quantified using a CPE potency assay, referenced against IFN α-2a reference standard, which has been tested and calibrated in International Units against the WHO reference standard. The three-dimensional structure of the present recombinant interferon (SEQ ID NO: 3, without N-terminal M) is different from the three-dimensional structure of IFN α-2b (SEQ ID NO: 4) published in the art (see Figure 2) and the three-dimensional structure of
based on computational modeling. There are obvious differences between the AB loops of the two, and their BC loops also cannot overlap completely (see Figure 3) .
Stability was tested according to ICH guideline for biological products Q1A and Q5C. The stability of the rSIFN-co composition is tested under 4-8℃ for 6 months, -80℃ for 24 months, and accelerated condition at 25℃ for 1 month. Repeated freeze/thaw cycle tests are done for 7 cycles. Stability data for 2 representative batches of drug substance 201801003, and 201801004 is shown in the following Tables 7-10.
Table 7. Stability Data of Long-term Test for Drug Substance of 2018 Batches (Y201801003 and Y201801004) at 4-8℃
Table 8. Stability Data of Long-term Test for Drug Substance of 2018 Batches (Y201801003 and Y201801004) at -80℃
Table 9. Stability Data of Accelerated Test for Drug Substance of 2018 Batches (Y201801003 and Y201801004) at 25±2℃
Table 10. Stability Data of Repeated Freezing and Thawing Test for Drug Substance of 2018 Batches (Y201801003 and Y201801004)
Example 2: Composition of Drug Product and Preparation
A composition of rSIFN-co was prepared by obtaining a 80%of the total volume of water for injection, adding a prescribed amount of disodium edetate, polysorbate 80, citric acid, glycerol, sodium chloride and benzyl alcohol in turn while stirring. Add the next component after the former component is fully dissolved. After each component is added, stir for more than 60 minutes until well combined. Cool the solution to below 20℃ and adjust pH value to about 5.0 with diluted hydrochloric acid. Stir the solution fully, add the prescribed amount of human serum albumin and rSIFN-co protein stock solution and then test pH value. If pH deviates, adjust pH value again to 5.0 and stir slightly. Dilute with water for injection cooled to below 20℃ to volume, homogenizing by gently stirring for more than 30 minutes before filter and sterilize with a 0.22μm filter to obtain the semi-finished rSIFN-co spray. A typical workflow is shown in Figure 5.
The current stability program and data showed that the composition (alternatively “drug product” ) is stable under 4-8℃ for 3 months and the real time study is on-going. It is also stable under accelerated condition at 25℃ for 2 months and at 37℃ for 2 weeks.
In one embodiment, the main components of the drug product are shown in the Table 11.
Table 11. Representative Formulation
In one embodiment, the product is a homogeneous colorless or light yellow with slightly sticky liquid spray. In one embodiment, the product is filled in high-density polyethylene bottles for pharmaceuticals with a mechanical nasal spray pump. Each bottle contains 8 ml and each pump is 0.1 ml.
In one embodiment, a solution is prepared according to the formula followed by sterilization and filtering by a 0.22μm filter. The filtered liquid was transferred into a sterile container to obtain a semi-finished product. After the sterility of the filtered liquid sample and the integrity of the used filter are tested, the sterile filtered solution is packaged into 10 ml medicinal high-density polyethylene bottle for external use by filling machine and equip with medicinal spray pump.
The final product is stored at a controlled temperature of between 2-8℃, and the latest stability data showed that the drug product is stable under 4-8℃ for 3 months and at accelerated condition of 25℃ for 2 months and at 37℃ for 2 weeks. Based on these test results at this time, the data supports a shelf life of 12 months and the stability program is on-going to support longer shelf life.
The drug product is supplied in spray bottle with mechanical pump. The components of the container closure system are in conformance to the compendial requirements for spray bottle containers for pharmaceutical use. The drug product is produced in GMP qualified facilities under ISO5, ISO7, and ISO8 clean room classification.
The detailed specification of the drug product is in Table 12.
Table 12. Drug Product Specification
Three batches of drug product have been manufactured to-date with batch number 20200209, 20200210 and 20200311. The test results of these three batches are shown as follows.
Table 13. Test Results for 3 Batches of Drug Product
Stability was established according to ICH guideline for biological products Q1A and Q5C. Sponsor plans to test the drug product stability under 4-8℃ for 36 months, at accelerated conditions 25℃ for 6 months and 37℃ for 1 month. Stability data for 3 representative batches of drug product 20200209, 20200210, and 20200311 is shown in the following Tables 14-17.
Table 14. Stability Data of Long-term Test for Spray Drug Product of 2020 Batches (20200209, 20200210 and 20200311) at 4~8℃
Table 15. Stability Data of Accelerated Test for Spray Drug Product of 2020 Batches (20200209, 20200210 and 20200311) at 25±2℃
Table 16. Stability Data of Accelerated Test for Spray Drug Product of 2020 Batches (20200209, 20200210 and 20200311) at 37±2℃
Example 3: Treatment by rSIFN-co of COVID-19 patients with symptoms
In this example, COVID-19 patients with symptoms such as mild pneumonia are treated by rSIFN-co or placebo for up to 10 to 28 days. rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time. Clinical improvement is determined by assessing radiological improvement and/or virus nucleic acid negative conversion at the end of treatment period. The median time to reach clinical improvement is approximately 8 to 16 days, and an overall rate of clinical improvement at the end of treatment is approximately 80%to 90%.
Example 4: Treatment by rSIFN-co of COVID-19 patients without symptoms
In this example, COVID-19 patients without symptoms are treated by rSIFN-co or placebo for up to 10 to 28 days. rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time. Any COVID-19 symptoms are assessed during the course of treatment. Patients treated with rSIFN-co are expected to show no symptom throughout the study, or significantly less treated patients show symptoms compared to patients receiving placebo.
Example 5: Treatment by rSIFN-co in combination with other agents of COVID-19 patients with symptoms
In this example, COVID-19 patients with symptoms such as mild pneumonia are treated by rSIFN-co in the presence of one or more agents for up to 10 to 28 days. rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time. During the treatment period, one or more of the following agents are also administered: (1) lopinavir (twice a day, 200 mg each time) , (2) ritonavir (twice a day, 50 mg each time) , (3) arbidol or umifenovir (three times a day, 200 mg each time) , (4) Remdesivir (100 mg per day) , (5) hydroxychloroquine (200 mg per day) , and (6) dexamethasone (4 mg per day) . Clinical improvement is determined by assessing radiological improvement and/or virus nucleic acid negative conversion at the end of treatment period. The median time to reach clinical improvement and virus nucleic acid negative conversion is expected to be significantly shorter in patients receiving rSIFN-co than patients not receiving rSIFN-co.
Example 6: Safety and tolerability in healthy subjects receiving rSIFN-co
In this example, subjects receive rSIFN-co by nebulization or injection, or placebo for up to 10 to 28 days. Any symptoms or adverse events are assessed and recorded during the course of the study.
Example 7: Treatment by rSIFN-co of COVID-19 patients on respirators
In this example, COVID-19 patients on respirators are treated by rSIFN-co or placebo for up to 10 to 28 days. rSIFN-co is administered by nebulization or injection twice a day, 8 to 12 million International Units (IU) each time. Clinical improvement is determined by assessing blood oxygen level during and at the end of treatment period. The median time to reach clinical improvement is approximately 8 to 16 days, and an overall rate of clinical improvement at the end of treatment is approximately 80%to 90%.
Example 8: Treatment by rSIFN-co Spray
In this example, healthy subjects or patients with an asymptomatic infection of Covid-19 receive 2 sprays per nostril and 4 sprays through pharynx once daily. Each spray comprises 2 μg of rSIFN-co. The period of treatment is approximately 5 days to 20 days. Subjects treated with rSIFN-co are expected to show no symptom throughout the study, or significantly less treated patients show symptoms compared to patients receiving placebo.
As for patients with mild or severe infection of Covid-19 receive, they get 2 sprays per nostril and 4 sprays through pharynx twice daily. Each spray comprises 2 μg of rSIFN-co. The period of treatment is approximately 10 days to 30 days. The median time to reach clinical improvement and virus nucleic acid negative conversion is expected to be significantly shorter in patients receiving rSIFN-co than patients not receiving rSIFN-co.
REFERENCES
1. WHO. Coronavirus disease (COVID-19) Weekly Epidemiological Update. 2020.
2. Wu, Z.; McGoogan, J.M. Characteristics of and Important Lessons From the Coronavirus Disease 2019 (COVID-19) Outbreak in China: Summary of a Report of 72 314 Cases From the Chinese Center for Disease Control and Prevention. Jama 2020, 10.1001/jama. 2020.2648, doi: 10.1001/jama. 2020.2648.
3. WHO. Clinical management of COVID-19.2020.
4. Stokes, E.K.; Zambrano, L.D.; Anderson, K.N.; Marder, E.P.; Raz, K.M.; Felix, S.E.B.; Tie, Y.; Fullerton, K.E. Coronavirus disease 2019 case surveillance-United States, January 22–May 30, 2020. Morbidity and Mortality Weekly Report 2020.
5. Huang, C.; Wang, Y.; Li, X.; Ren, L.; Zhao, J.; Hu, Y.; Zhang, L.; Fan, G.; Xu, J.; Gu, X., et al. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China. Lancet 2020, 395, 497-506, doi: 10.1016/s0140-6736 (20) 30183-5.
6. Chan, J.F.; Kok, K.H.; Zhu, Z.; Chu, H.; To, K.K.; Yuan, S.; Yuen, K.Y. Genomic characterization of the 2019 novel human-pathogenic coronavirus isolated from a patient with atypical pneumonia after visiting Wuhan. Emerg Microbes Infect 2020, 9, 221-236, doi: 10.1080/22221751.2020.1719902.
7. Letko, M.; Marzi, A.; Munster, V. Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses. Nat Microbiol 2020, 5, 562-569, doi: 10.1038/s41564-020-0688-y.
8. Karia, R.; Gupta, I.; Khandait, H.; Yadav, A.; Yadav, A. COVID-19 and its Modes of Transmission. SN Compr Clin Med 2020, 10.1007/s42399-020-00498-4, 1-4, doi: 10.1007/s42399-020-00498-4.
9. China, N.H.C.o.t.P.s.R.o. Diagnosis and treatment of pneumonia caused by new coronavirus infection (version 8) . Beijing, China. Availabe online: www. nhc. gov. cn/yzygj/s7653p/202008/0a7bdf12bd4b46e5bd28ca7f9a7f5e5a. shtml (accessed on 16 September) .
10. Wei, W.E.; Li, Z.; Chiew, C.J.; Yong, S.E.; Toh, M.P.; Lee, V.J. Presymptomatic Transmission of SARS-CoV-2 -Singapore, January 23–March 16, 2020. Morb Mortal Wkly Rep 2020, 69, 5.
11. Tong, Z.D.; Tang, A.; Li, K.F.; Li, P.; Wang, H.L.; Yi, J.P.; Zhang, Y.L.; Yan, J.B. Potential Presymptomatic Transmission of SARS-CoV-2, Zhejiang Province, China, 2020. Emerg Infect Dis 2020, 26, 1052-1054, doi: 10.3201/eid2605.200198.
12. van Kampen, J.J.A.; van de Vijver, D.A.M.C.; Fraaij, P.L.A.; Haagmans, B.L.; Lamers, M.M.; Okba, N.; van den Akker, J.P.C.; Endeman, H.; Gommers, D.A.M.P.J.; Cornelissen, J.J., et al. Shedding of infectious virus in hospitalized patients with coronavirus disease-2019 (COVID-19) : duration and key determinants. medRxiv 2020, 10.1101/2020.06.08.20125310, 2020.2006.2008.20125310, doi: 10.1101/2020.06.08.20125310.
13. La Scola, B.; Le Bideau, M.; Andreani, J.; Hoang, V.T.; Grimaldier, C.; Colson, P.; Gautret, P.; Raoult, D. Viral RNA load as determined by cell culture as a management tool for discharge of SARS-CoV-2 patients from infectious disease wards. Eur J Clin Microbiol Infect Dis 2020, 39, 1059-1061, doi: 10.1007/s10096-020-03913-9.
14. He, X.; Lau, E.H.Y.; Wu, P.; Deng, X.; Wang, J.; Hao, X.; Lau, Y.C.; Wong, J.Y.; Guan, Y.; Tan, X., et al. Temporal dynamics in viral shedding and transmissibility of COVID-19. Nat Med 2020, 26, 672-675, doi: 10.1038/s41591-020-0869-5.
15. Zhang, K.J.; Yin, X.F.; Yang, Y.Q.; Li, H.L.; Xu, Y.N.; Chen, L.Y.; Liu, X.J.; Yuan, S.J.; Fang, X.L.; Xiao, J., et al. A Potent In Vivo Antitumor Efficacy of Novel Recombinant Type I Interferon. Clin Cancer Res 2017, 23, 2038-2049, doi: 10.1158/1078-0432. Ccr-16-1386.
16. Samuel, C.E. Antiviral actions of interferons. Clin Microbiol Rev 2001, 14, 778-809, table of contents, doi: 10.1128/cmr. 14.4.778-809.2001.
17. Hadjadj, J.; Yatim, N.; Barnabei, L.; Corneau, A.; Boussier, J.; Smith, N.; Péré, H.; Charbit, B.; Bondet, V.; Chenevier-Gobeaux, C., et al. Impaired type I interferon activity and inflammatory responses in severe COVID-19 patients. Science 2020, 369, 718-724, doi: 10.1126/science. abc6027.
18. Acharya, D.; Liu, G.; Gack, M.U. Dysregulation of type I interferon responses in COVID-19. Nat Rev Immunol 2020, 20, 397-398, doi: 10.1038/s41577-020-0346-x.
19. Martinez, M.A. Compounds with Therapeutic Potential against Novel Respiratory 2019 Coronavirus. Antimicrobial Agents and Chemotherapy 2020, 64, e00399-00320, doi: 10.1128/aac. 00399-20.
20. Sallard, E.; Lescure, F.X.; Yazdanpanah, Y.; Mentre, F.; Peiffer-Smadja, N. Type 1 interferons as a potential treatment against COVID-19. Antiviral Res 2020, 178, 104791, doi: 10.1016/j. antiviral. 2020.104791.
21. Meng, Z.; Wang, T.; Chen, L.; Chen, X.; Li, L.; Qin, X.; Li, H.; Luo, J. An experimental trial of recombinant human interferon alpha nasal drops to prevent COVID-19 in medical staff in an epidemic area. medRxiv 2020, 10.1101/2020.04.11.20061473, doi: 10.1101/2020.04.11.20061473.
22. Davoudi-Monfared, E.; Rahmani, H.; Khalili, H.; Hajiabdolbaghi, M.; Salehi, M.; Abbasian, L.; Kazemzadeh, H.; Yekaninejad, M.S. A Randomized Clinical Trial of the Efficacy and Safety of Interferon β-1a in Treatment of Severe COVID-19. Antimicrobial Agents and Chemotherapy 2020, 64, e01061-01020, doi: 10.1128/aac. 01061-20.
23. Identifier: ChiCTR2000029638. A Multicenter, Randomized, Controlled trial for Recombinant Super-Compound Interferon (rSIFN-co) in the Treatment of 2019 Novel Coronavirus (2019-nCoV) Infected Pneumonia. 2020. See also Chinese patent application publication 111658779A.
24. Chen, Q.; Zhang, L.L.; Yu, D.X.; Yu, Z.A.; Liu, Y.; Zhang, L.P.; Li, Z.F.; Duan, Z.J.; Wang, B.H.; Wei, X.J., et al. [A field trial for evaluating the safety of recombinant human interferon alpha-2b for nasal spray] . Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi 2005, 19, 211-215.
25. Panel, C. -T.G. Coronavirus Disease 2019 (COVID-19) Treatment Guidelines. Availabe online: https: //www. covid19treatmentguidelines. nih. gov/ (accessed on 17 September) .
26. CDC. Centers for Disease Control and Prevention. Interim guidelines for collecting, handling, and testing clinical specimens from persons for coronavirus disease 2019 (COVID-19) . Availabe online: https: //www. cdc. gov/coronavirus/2019-ncov/lab/guidelines-clinical-specimens. html (accessed on Sep 14) .
27. A minimal common outcome measure set for COVID-19 clinical research. Lancet Infect Dis 2020, 20, e192-e197, doi: 10.1016/s1473-3099 (20) 30483-7.
.
Claims (17)
- A method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject, said method comprising administering to the subject a therapeutically effective dose of recombinant super-compound interferon (rSIFN-co) for a period of at least 3 days.
- The method of claim 1, wherein said rSIFN-co is administered to said subject by nasal spray, pharynx spray or intravenous injection.
- The method of claim 1, wherein said therapeutically effective dose ranges from 5 μg to 100 μg per day.
- The method of claim 1, wherein said therapeutically effective dose is selected from the group consisting of 8 μg, 16 μg, 32 μg, 64 μg and 96 μg per day.
- The method of claim 1, wherein said period is 5 to 30 days.
- The method of claim 1, wherein said subject suffers from one or more diseases in one or more of the digestive system, liver, heart, and kidney.
- The method of claim 1, wherein said subject receives one or more additional treatments selected from the group consisting of Remdesivir, hydroxychloroquine, and dexamethasone.
- The method of claim 2, wherein said subject is healthy or has an asymptomatic infection of Covid-19 coronavirus, and said subject receives 2 sprays per nostril and 4 sprays through pharynx once daily, each spray comprising 0.2-5.0 μg of said rSIFN-co.
- The method of claim 2, wherein said subject shows mild covid-19 infection and receives 2 sprays per nostril and 4 sprays through pharynx twice daily, each spray comprising 0.2-5.0 μg of said rSIFN-co.
- The method of claim 1, wherein the rSIFN-co has a specific activity greater than 5.0 x 10 8 IU/mg protein.
- The method of claim 2, wherein the nasal spray, pharynx spray or intravenous injection comprises one or more of stabilizer, bacteriostat, surfactant, metal complex, isosmotic adjusting agent, buffering agent, and viscosity modifier.
- The method of claim 12, wherein said stabilizer is human serum albumin, the metal complex is disodium edetate, the surfactant is Tween 80, the buffering agent is citric acid, the viscosity modifier is glycerol, the isosmotic adjusting agent is sodium chloride, and the bacteriostat is benzyl alcohol.
- The method of claim 2, wherein the nasal spray, pharynx spray or intravenous injection has a pH value of about 5.0.
- The method of claim 2, wherein the nasal spray or pharynx spray is stable under 4-8℃ for 3 months, at 25℃ for 2 months and at 37℃ for 2 weeks.
- A method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject, said method comprising administering to the subject a composition comprising a therapeutically effective dose of rSIFN-co for a period of at least 3 days.
- A method of preventing or treating Covid-19 (SARS-Cov-2) coronavirus infection in a subject, said method comprising administering to the subject a composition consisting of a therapeutically effective dose of rSIFN-co for a period of at least 3 days.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US202163145496P | 2021-02-04 | 2021-02-04 | |
US63/145,496 | 2021-02-04 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2022166885A1 true WO2022166885A1 (en) | 2022-08-11 |
Family
ID=82740853
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/CN2022/074977 WO2022166885A1 (en) | 2021-02-04 | 2022-01-29 | Recombinant super-compound interferon (rsifn-co) for treating covid-19 patients with or without symptoms |
Country Status (1)
Country | Link |
---|---|
WO (1) | WO2022166885A1 (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2011202683A1 (en) * | 2003-08-28 | 2011-06-30 | Superlab Far East Limited | Uses of interferons with altered spatial structure |
CN111346219A (en) * | 2020-02-21 | 2020-06-30 | 上海甘翼生物医药科技有限公司 | Use of interferon in preparing medicine for preventing coronavirus infection or preventing diseases caused by coronavirus infection |
CN111658779A (en) * | 2020-06-22 | 2020-09-15 | 四川大学华西医院 | Combined medicine for treating novel coronavirus pneumonia |
-
2022
- 2022-01-29 WO PCT/CN2022/074977 patent/WO2022166885A1/en active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2011202683A1 (en) * | 2003-08-28 | 2011-06-30 | Superlab Far East Limited | Uses of interferons with altered spatial structure |
CN111346219A (en) * | 2020-02-21 | 2020-06-30 | 上海甘翼生物医药科技有限公司 | Use of interferon in preparing medicine for preventing coronavirus infection or preventing diseases caused by coronavirus infection |
CN111658779A (en) * | 2020-06-22 | 2020-09-15 | 四川大学华西医院 | Combined medicine for treating novel coronavirus pneumonia |
Non-Patent Citations (2)
Title |
---|
FRAGKOU P.C., BELHADI D., PEIFFER-SMADJA N., MOSCHOPOULOS C.D., LESCURE F.-X., JANOCHA H., KAROFYLAKIS E., YAZDANPANAH Y., MENTRÉ : "Review of trials currently testing treatment and prevention of COVID-19", CLINICAL MICROBIOLOGY AND INFECTION., WILEY-BLACKWELL PUBLISHING LTD, UNITED KINGDOM, SWITZERLAND, vol. 26, no. 8, 1 August 2020 (2020-08-01), United Kingdom, Switzerland , pages 988 - 998, XP055955419, ISSN: 1198-743X, DOI: 10.1016/j.cmi.2020.05.019 * |
PEREDA RICARDO, GONZÁLEZ D, RIVERO HB, RIVERO JC, PÉREZ A, LÓPEZ LR, MEZQUIA N, VENEGAS R, BETANCOURT JULIO RAUL, DOMÍNGUEZ RE: "Therapeutic effectiveness of interferon alpha 2b treatment for COVID-19 patient recovery", MEDRXIV, 4 August 2020 (2020-08-04), XP055955420, Retrieved from the Internet <URL:https://www.medrxiv.org/content/medrxiv/early/2020/08/04/2020.07.28.20157974.full.pdf> DOI: 10.1101/2020.07.28.20157974 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Xu et al. | Arbidol/IFN-α2b therapy for patients with corona virus disease 2019: a retrospective multicenter cohort study | |
De Luna et al. | Rapid and severe Covid‐19 pneumonia with severe acute chest syndrome in a sickle cell patient successfully treated with tocilizumab | |
US20070134763A1 (en) | Anti-virus therapy for respiratory diseases | |
JP2002542202A (en) | HCV combination therapeutic containing ribavirin with antioxidant | |
US11364227B2 (en) | Sphingosine kinase 2 inhibitor for treating coronavirus infection | |
EP1922081B1 (en) | Interferon lambda therapy for treatment of respiratory diseases | |
WO2021164740A1 (en) | Use of interferon in preparing drug for preventing coronavirus infection or preventing disease caused by coronavirus infection | |
CN111671886B (en) | Pharmaceutical composition for preventing high-risk susceptible people from infecting coronavirus or generating coronavirus infection disease and application of pharmaceutical composition | |
WO2022166885A1 (en) | Recombinant super-compound interferon (rsifn-co) for treating covid-19 patients with or without symptoms | |
Murry et al. | Respiratory syncytial virus: not just for kids | |
Adams Jr et al. | Interferon treatment of adenoviral conjunctivitis | |
US11278598B2 (en) | Methods for treating diseases associated with respiratory viruses | |
Garcia-Huidobro et al. | Safety, tolerability, bioavailability, and biological activity of inhaled interferon-α2b in healthy adults: the IN2COVID Phase I randomized trial | |
US12115150B2 (en) | Biomarkers of coronavirus pneumonia | |
US20230241177A1 (en) | Use of inhaled interferon-beta to treat virus-induced exacerbations in copd patients undergoing treatment with a systemic corticosteroid | |
JP7579880B2 (en) | Application of the combination of TFF2 protein and IFN-κ protein in the treatment of novel coronavirus infection | |
US20230338475A1 (en) | USE OF INHALED INTERFERON-BETA TO IMPROVE OUTCOME IN SARS-CoV-2 INFECTED PATIENTS | |
Nair et al. | Immunity Modulators, Repurposed Drugs and Candidate Vaccines for COVID-19: A Review | |
US20230201308A1 (en) | Treatment of coronavirus infection with interferon lambda | |
Rodrigues et al. | A brief overview of current drug repurposing approaches for COVID-19 management | |
KR20220139922A (en) | Treatment of coronavirus infection with interferon lambda | |
CN116472054A (en) | Inhaled interferon-beta for improving prognosis of SARS-CoV-2 infected patient | |
Chen et al. | Severe Acute Respiratory Syndrome Coronavirus 2 Infection in Patients Living with Human Immunodeficiency Virus: Case Reports and Review of the Literature | |
WO2024038186A1 (en) | Treatment of acute respiratory failure | |
Kjellberg et al. | Ten Sessions of Hyperbaric Oxygen Versus Sham Treatment in Patients with Long COVID (HOT-LoCO): A Randomised, Placebo Controlled, Double-Blind, Phase II Trial |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 22749164 Country of ref document: EP Kind code of ref document: A1 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
32PN | Ep: public notification in the ep bulletin as address of the adressee cannot be established |
Free format text: NOTING OF LOSS OF RIGHTS PURSUANT TO RULE 112(1) EPC (EPO FORM 1205A DATED 17.01.2024) |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 22749164 Country of ref document: EP Kind code of ref document: A1 |