WO2021238982A1 - 包含聚核苷酸的药物组合物及其用于预防或治疗covid-19的用途 - Google Patents

包含聚核苷酸的药物组合物及其用于预防或治疗covid-19的用途 Download PDF

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Publication number
WO2021238982A1
WO2021238982A1 PCT/CN2021/096048 CN2021096048W WO2021238982A1 WO 2021238982 A1 WO2021238982 A1 WO 2021238982A1 CN 2021096048 W CN2021096048 W CN 2021096048W WO 2021238982 A1 WO2021238982 A1 WO 2021238982A1
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Prior art keywords
unit dose
unit
active
cov
dose
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PCT/CN2021/096048
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English (en)
French (fr)
Inventor
张译
刘媛
张楠
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beijing Yisheng Xingye Technology Co Ltd
Henan Yisheng Pharma Co Ltd
Liaoning Yisheng Biopharma Co Ltd
Original Assignee
Beijing Yisheng Xingye Technology Co Ltd
Henan Yisheng Pharma Co Ltd
Liaoning Yisheng Biopharma Co Ltd
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Priority to CA3178162A priority Critical patent/CA3178162A1/en
Priority to IL298611A priority patent/IL298611A/en
Priority to JP2023516641A priority patent/JP2023540643A/ja
Priority to BR112022024186A priority patent/BR112022024186A2/pt
Priority to AU2021278137A priority patent/AU2021278137A1/en
Priority to MX2022014676A priority patent/MX2022014676A/es
Priority to KR1020227045011A priority patent/KR20230013133A/ko
Priority to US17/768,441 priority patent/US20230081039A1/en
Application filed by Beijing Yisheng Xingye Technology Co Ltd, Henan Yisheng Pharma Co Ltd, Liaoning Yisheng Biopharma Co Ltd filed Critical Beijing Yisheng Xingye Technology Co Ltd
Priority to CU2022000068A priority patent/CU20220068A7/es
Priority to EP21812811.4A priority patent/EP4159233A4/en
Priority to CN202180004682.1A priority patent/CN114340662A/zh
Publication of WO2021238982A1 publication Critical patent/WO2021238982A1/zh
Priority to ZA2022/12363A priority patent/ZA202212363B/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/215—Coronaviridae, e.g. avian infectious bronchitis virus
    • 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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • 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
    • A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/7036—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin having at least one amino group directly attached to the carbocyclic ring, e.g. streptomycin, gentamycin, amikacin, validamycin, fortimicins
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
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    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
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    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
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    • A61K39/00—Medicinal preparations containing antigens or antibodies
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    • A61K2039/55561—CpG containing adjuvants; Oligonucleotide containing adjuvants
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    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/20011—Coronaviridae
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    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/20011—Coronaviridae
    • C12N2770/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/20011—Coronaviridae
    • C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • This application relates to the medical field, especially the prevention or treatment of coronavirus infection. More specifically, it relates to a pharmaceutical composition for preventing or treating coronavirus infection, which contains: polyriboinosine-polyribocytidylic acid (PIC), antibiotics (or polyamino compounds), positive ions, And optionally antigens derived from viruses (for example, but not limited to: virus particles, proteins, nucleic acids, peptides); and the use of the composition for preventing or treating coronavirus infections.
  • PIC polyriboinosine-polyribocytidylic acid
  • antibiotics or polyamino compounds
  • positive ions for example, but not limited to: virus particles, proteins, nucleic acids, peptides
  • antigens derived from viruses for example, but not limited to: virus particles, proteins, nucleic acids, peptides
  • the new type of coronavirus infection is mainly caused by the new type of coronavirus (SARS-CoV-2, also known as 2019-nCOV).
  • SARS-CoV-2 also known as 2019-nCOV.
  • the main route of infection is through respiratory droplets, but also through contact.
  • the population is generally susceptible, and the elderly and those with underlying diseases are more ill after infection, and children and infants also have the disease.
  • the incubation period of COVID-19 is 1-14 days, mostly 3-7 days.
  • the main manifestations are fever, dry cough, and fatigue.
  • a small number of patients are accompanied by nasal congestion, runny nose, sore throat, myalgia, and diarrhea.
  • Severe patients often develop dyspnea and/or hypoxemia one week after the onset of onset. In severe cases, they can rapidly progress to acute respiratory distress syndrome, septic shock, difficult to correct metabolic acidosis, coagulation dysfunction, and multiple organs Exhaustion and so on. It is worth noting that severe and critically ill patients may have moderate to low fever during the course of their illness, or even no obvious fever.
  • the present disclosure provides an immune composition comprising polyriboinosine-polyribocytidylic acid (polyI:C, PIC), antibiotics (or polyamino compounds), positive ions and optionally virus-derived antigens ; Or, it is composed of PIC, antibiotics (or polyamino compounds), positive ions and optional viral antigens.
  • the virus is a coronavirus, such as SARS-CoV-2 (including its variants or subspecies).
  • the virus is inactivated crude or inactivated purified type.
  • vaccines or immune compositions suitable for the present disclosure include, but are not limited to, inactivated type, subunit type, genetically recombined type, polypeptide type vaccine, and nucleic acid type.
  • the PIC suitable for the present application may be a commercially available PIC, or a polyinosinic acid-polycytidylic acid-based adjuvant disclosed in the art (for example, 2006800007767 and 2006800010596), which are incorporated by reference in their entirety. Enter here.
  • the PIC in the immune composition is heterogeneous in molecular weight, with a molecular weight of at least 50,000 Daltons.
  • a value of 66,000 Daltons corresponds to a molecular size in 6.4S sedimentation coefficient units (Svedbergs).
  • the PIC is 66,000 Daltons to 1,200,000 Daltons (equivalent to 6.4 to 24.0 sedimentation coefficient units).
  • the molecular weight of the PIC is at least 150,000 Daltons.
  • the molecular weight of the PIC is 100,000 to 200,000 Daltons, or 300,000 to 4,000,000 Daltons, or 500,000 to 1,000,000 Daltons, or 1,000,000 to 1,500,000 Daltons, or 1,500,000 to 2,000,000 Daltons Daltons, or 2,000,000 to 2,500,000 Daltons, or 2,500,000 to 3,000,000 Daltons, or 3,000,000 to 3,500,000 Daltons, or 3,500,000 to 4,000,000 Daltons, or 4,000,000 to 4,500,000 Daltons, or It is 4,500,000 to 5,000,000 Daltons.
  • heterogeneous as used in the present disclosure means that the PIC molecules contained in the immune composition or vaccine are not uniform in molecular weight, size, or physical properties of both.
  • the concentration of the PIC in the immune composition/vaccine is 250 ⁇ g/unit dose to 5000 ⁇ g/unit dose; for example, the concentration of the PIC in the composition is selected from: 250 ⁇ g/unit dose, 500 ⁇ g/unit dose , 1000 ⁇ g/unit dose, 1500 ⁇ g/unit dose, 2000 ⁇ g/unit dose, 3000 ⁇ g/unit dose, 4000 ⁇ g/unit dose, 5000 ⁇ g/unit dose, and the range between any two points above.
  • the concentration of PIC in the immune composition/vaccine is 500 ⁇ g/unit dose to 4000 ⁇ g/unit dose, 1000 ⁇ g/unit dose to 3000 ⁇ g/unit dose, or 1000 ⁇ g/unit dose to 2500 ⁇ g/unit dose.
  • the concentration of PIC in the immune composition/vaccine is selected from 500 ⁇ g/unit dose, 1000 ⁇ g/unit dose, 1500 ⁇ g/unit dose, 2000 ⁇ g/unit dose, and any two of the above. The range between.
  • the concentration of PIC in the immune composition/vaccine is selected from 250 ⁇ g/unit dose, 500 ⁇ g/unit dose, 1000 ⁇ g/unit dose, 1250 ⁇ g/unit dose And the range between any two points above.
  • the antibiotics contained are selected from the group consisting of: tetracycline, anthracycline, butyricin sulfate, gentamicin, hygromycin, amikonin, kanamycin Dideoxykanamycin (or dideoxykanamycin), cyanomycin, metatamycin, neomycin, puromycin, streptomycin, streptozotocin, and combinations thereof.
  • the polyamino compound is selected from the group consisting of: arginine salt, quinine, N-(3-aminopropyl), N-(3-aminopropyl)-1,4-butanediamine, spermine , Sperm, OS-dimethylamino phosphorothioate, polylysine, aminoglycoside and combinations thereof.
  • the concentration of the antibiotic in the composition is 10 units/ml to 100,000 units/ml, preferably 100 units/ml to 10,000 units/ml, more preferably 500 units/ml to 5,000 units/ml.
  • the concentration of antibiotics or polyamino compounds in the immune composition can be mentioned from 400 U/unit dose to 1200 U/unit dose, such as but not limited to 400, 500, 600, 700, 800, 900, 1000 , 1100, 1200U/unit dose.
  • the concentration of antibiotics or polyamino compounds in the immune composition can be mentioned to be 400U/ml to 1200U/ml, such as but not limited to 400, 500, 600, 700, 800, 900, 1000, 1100, 1200U/ml.
  • the positive ions contained are divalent positive ions, and are selected from calcium, cadmium, lithium, magnesium, cerium, cesium, chromium, cobalt, deuterium, gallium, iodine, iron, zinc, and combination.
  • the positive ion is calcium ion.
  • the positive ion may be in the form of any suitable salt or organic complex, including but not limited to: chloride, fluoride, hydroxide, phosphate, or sulfate.
  • the positive ion when the positive ion is calcium, the calcium ion may be in the form of calcium carbonate, calcium chloride, calcium fluoride, calcium hydroxide, calcium phosphate, or calcium sulfate.
  • the concentration of the positive ion in the composition is 10 ⁇ mol to 10 mmol/ml, for example, 50 ⁇ mol to 5 mmol/ml; or 100 ⁇ mol to 1 mmol/ml.
  • the concentration of the positive ion in the immune composition is 0.01 mg/unit dose to 0.1 mg/unit dose, such as but not limited to 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 , 0.1mg/unit dose.
  • the concentration of the positive ion in the immune composition is 0.01 mg/ml to 0.1 mg/ml, such as but not limited to 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1mg/ml.
  • the coronavirus-derived antigen is a virus (e.g., attenuated, inactivated virus, artificially assembled viral vector or viral particle).
  • the coronavirus-derived antigen is the inactivated virus SARS-CoV-2, the attenuated virus SARS-CoV-2, or the virus SARS-CoV-2 that cannot reproduce in the subject.
  • the ratio of the virus to the PIC is selected from: 1 activity unit/50 ⁇ g, 1 activity unit/60 ⁇ g, 1 activity unit/70 ⁇ g, 1 activity unit/80 ⁇ g, 1 activity unit/90 ⁇ g, 1 activity unit/100 ⁇ g, 1 activity unit /125 ⁇ g, 1 active unit/200 ⁇ g, 1 active unit/250 ⁇ g, 1 active unit/300 ⁇ g, 1 active unit/350 ⁇ g, 1 active unit/400 ⁇ g, 1 active unit/450 ⁇ g, 1 active unit/500 ⁇ g, 1 active unit/550 ⁇ g , 1 active unit/600 ⁇ g, 1 active unit/700 ⁇ g, 1 active unit/800 ⁇ g, 1 active unit/1000 ⁇ g, 1 active unit/1500 ⁇ g, 1 active unit/2000 ⁇ g, 1 active unit/2500 ⁇ g,
  • the concentration of the antigen in the composition is 0.2 active unit/ Unit dose to 100 active units/unit dose.
  • the concentration of the virus or virus-derived antigen in the composition is selected from 0.2 active unit/unit dose, 0.5 active unit/unit dose, 1.0 active unit/unit dose, 1.5 active unit/unit dose, 2.0 Active unit/unit dose, 2.5 active unit/unit dose, 3.0 active unit/unit dose, 3.5 active unit/unit dose, 4.0 active unit/unit dose, 5.0 active unit/unit dose, 6.0 active unit/unit dose, 7.0 active Unit/unit dose, 8.0 active unit/unit dose, 9.0 active unit/unit dose, 10.0 active unit/unit dose, 15.0 active unit/unit dose, 20.0 active unit/unit dose, 30.0 active unit/unit dose, 40.0 active unit /Unit dose, 50.0 active unit/unit dose, 60.0 active unit/unit dose, 70.0 active unit/unit dose, 80.0 active unit/unit dose, 90.0 active unit/unit dose, 100.0 active unit/unit dose, and any two of the above The range between points.
  • the concentration of the virus or virus-derived antigen in the composition is 0.5 active unit/unit dose, 1.0
  • the concentration of the antigen in the composition is 0.05 active unit/ ml to 40.0 active unit/ml; preferably 0.05 active unit/ml, 0.1 active unit/ml, 0.15 active unit/ml, 0.2 active unit/ml, 0.5 active unit/ml, 1.0 active unit/ml, 2.0 active unit/ml , 3.0 active units/ml, 4.0 active units/ml, 5.0 active units/ml, 10 active units/ml, 15 active units/ml, 20 active units/ml, 25 active units/ml, 30 active units/ml, 35 Activity unit/ml, 40 activity unit/ml.
  • the concentration of the antigen in the composition is 0.05 active unit/ ml to 40.0 active unit/ml; preferably 0.05 active unit/ml, 0.1 active unit/ml, 0.15 active unit/ml, 0.2 active unit/ml, 0.5 active unit/ml, 1.0 active unit/ml, 2.0 active unit/ml , 3.0 active units/ml,
  • the ratio of the antigen to the PIC is selected from: 1 ⁇ g/10 ⁇ g, 1 ⁇ g/20 ⁇ g, 1 ⁇ g/30 ⁇ g, 1 ⁇ g/40 ⁇ g, 1 ⁇ g/50 ⁇ g, 1 ⁇ g/60 ⁇ g, 1 ⁇ g/70 ⁇ g, 1 ⁇ g/80 ⁇ g, 1 ⁇ g/90 ⁇ g, 1 ⁇ g/100 ⁇ g, 1 ⁇ g/125 ⁇ g, 1 ⁇ g/200 ⁇ g, 1 ⁇ g/250 ⁇ g, 1 ⁇ g/300 ⁇ g, 1 ⁇ g/300 ⁇ g 350 ⁇ g, 1 ⁇ g/400 ⁇ g, 1 ⁇ g/450 ⁇ g, 1 ⁇ g/500 ⁇ g, 1 ⁇ g/550 ⁇ g, 1 ⁇ g/600
  • the coronavirus-derived antigen is selected from the following: S protein or immunogenic fragment thereof, M protein or immunogenic fragment thereof, N protein or immunogen thereof of SARS-CoV-2
  • concentration of sexual fragment, E protein or its immunogenic fragment, protein, polypeptide, DNA, RNA designed according to the SARS-CoV-2 structure in the immune composition is 0.1 ⁇ g/unit dose to 1000.0 ⁇ g/unit dose, For example but not limited to 0.5 ⁇ g/unit dose, 1.0 ⁇ g/unit dose, 2.0 ⁇ g/unit dose, 3.0 ⁇ g/unit dose, 4.0 ⁇ g/unit dose, 5.0 ⁇ g/unit dose, 6.0 ⁇ g/unit dose, 7.0 ⁇ g/unit Dose, 8.0 ⁇ g/unit dose, 9.0 ⁇ g/unit dose, 10.0 ⁇ g/unit dose, 15.0 ⁇ g/unit dose, 20.0 ⁇ g/unit dose, 30.0 ⁇ g/unit dose, 40.0 ⁇ g/unit dose, 50.0 ⁇ g/unit dose, 60.0 ⁇ g/unit dose, 70.0 ⁇ g/unit dose
  • the concentration of the antigen in the immune composition is 2.5 ⁇ g/unit dose to 20 ⁇ g/unit dose.
  • the concentration of the antigen in the immune composition is 2.5 ⁇ g/unit dose to 20 ⁇ g /Unit dose.
  • the concentration of the antigen in the immune composition is 2.5 ⁇ g/unit dose to 20 ⁇ g/unit dose.
  • the concentration of the antigen in the immune composition is 2.5 ⁇ g/unit dose to 20 ⁇ g/unit dose.
  • the unit dose described in the present disclosure is expressed in the form of volume, and is selected from: 0.1ml, 0.15ml, 0.2ml, 0.5ml, 1.0ml, 1.5ml, 2.0ml, 2.5ml, 3.0ml , 4.0ml, 5.0ml, 10.0ml, 20.0ml, 30.0ml, 40.0ml, 50.0ml, 60.0ml, 70.0ml, 80.0ml, 90.0ml, 100.0ml, 150ml, 200ml, 250.00ml, and any two of the above The range between.
  • the skilled person understands that too large or too small unit doses cause inconvenience in clinical operation.
  • the unit dose is preferably in the range of 0.5 ml to 1.0 ml.
  • the unit dose is preferably in the range of 0.15ml to 0.2ml.
  • the unit dose is preferably in the range of 30.0 ml to 1000 ml. It should be understood here that although the unit dose is expressed by volume, this does not mean that the immune composition/vaccine of the present disclosure can only be in liquid form.
  • the volume of a unit dose refers to the volume of the dry powder or lyophilized powder after reconstitution.
  • the immune composition or vaccine of the present disclosure may comprise gelatin, sucrose, white granulated sugar, lactose, maltose, trehalose, glucose, low molecular dextran, sorbitol, polysorbate 20, polysorbate Ester 80, arginine hydrochloride, mannitol polyethylene glycol, human albumin, recombinant albumin, sodium caprylate, urea, aluminum hydroxide, phenol red, aluminum phosphate, squalene, saponins, oligonucleotides, Magnesium chloride, potassium chloride, sodium chloride, sodium thiosulfate, potassium dihydrogen phosphate, ascorbic acid, chloroform, phenol, thimerosal.
  • the immune composition or vaccine of the present disclosure may also include a physiologically acceptable buffer selected from the group consisting of acetate, Tris, bicarbonate, carbonate, and phosphate buffer.
  • a physiologically acceptable buffer selected from the group consisting of acetate, Tris, bicarbonate, carbonate, and phosphate buffer.
  • the pH value of the buffer suitable for the composition of the present disclosure is selected from: 6.50, 6.60, 6.70, 6.80, 6.90, 7.00, 7.05, 7.1, 7.15, 7.2, 7.25, 7.30, 7.35, 7.40, 7.45, 7.50, 7.55, 7.60, 7.65, 7.70, 7.75, 7.80, 7.85, 7.90, 7.95, 8.00 and the range between any two points above.
  • the buffer is PBS, and the pH is in the range of 7.0 to 8.0.
  • the concentration of the buffer in the composition is 5 mM to 50 mM, specifically 5 mM to 20 mM. Examples that can be mentioned are 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50mM.
  • the pH of the buffer is not ruled out that its pH will change with the addition of other ingredients.
  • the pH value of the immune composition or vaccine of the present disclosure is controlled in an appropriate range, for example: 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8 , 6.9, 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35, 7.4, 7.45, 7.5, 7.55, 7.6, 7.65, 7.7, 7.75, 7.8, 7.85, 7.9, 7.95, 8.0, 8.1, 8.2, 8.3 , 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0 and the range between any two points above.
  • the pH of the composition is in the range of 7.0 to 8.0.
  • the immune composition or vaccine of the present disclosure can be prepared as dry powder, liquid solution (for example, injectable solution, aqueous or physiological saline solution, or suspension, ointment, droplet, emulsion, gel, syrup or slurry, gas Mist, spray, nasal drops), tablets, coated tablets, microcapsules, suppositories, tablets, granules, dragees, capsules.
  • liquid solution for example, injectable solution, aqueous or physiological saline solution, or suspension, ointment, droplet, emulsion, gel, syrup or slurry, gas Mist, spray, nasal drops
  • tablets coated tablets, microcapsules, suppositories, tablets, granules, dragees, capsules.
  • the immune composition or vaccine of the present disclosure is prepared as an injectable solution.
  • an immune composition which is in the form of a dry powder or a lyophilized powder corresponding to the above-mentioned immune composition.
  • the skilled person can understand that when the immune composition is in the form of a dry powder or a freeze-dried powder, the only difference from the liquid form of the immune composition is the water content.
  • the immune composition in the form of dry powder or lyophilized powder contains 2 mg/ml PIC, the skilled person knows that this concentration refers to the concentration of PIC after the immune composition is re-formulated into a liquid form.
  • an immune composition comprising:
  • an immune composition comprising:
  • the use of the combination of the above-mentioned PIC, antibiotic (or polyamino compound) and positive ion in the preparation of a medicine or vaccine for preventing or treating coronavirus infection is provided.
  • polynucleotide adjuvant composition disclosed in CN103405762A in the preparation of drugs or vaccines for preventing or treating coronavirus infections.
  • the use of the immune composition or vaccine of the present disclosure in preventing or treating coronavirus infection is provided.
  • the use of the immune composition or vaccine of the present disclosure in the preparation of a medicine for preventing or treating coronavirus infection is provided.
  • the present disclosure also relates to a method of stimulating an immune response in a host animal, the method comprising administering to the animal an amount of the immune composition described herein effective to induce an immune response.
  • the host animal is preferably a mammal, more preferably a monkey, still more preferably a human.
  • the present invention also relates to a method for immunizing a host animal against a virus, which method comprises administering to the animal an immune composition as described herein in an amount effective to induce a protective response.
  • the host animal is preferably a mammal, more preferably a monkey, and even more preferably a human.
  • an “immune response” to an antigen or composition is a response produced in the subject's humoral and/or cellular immune response against the molecules present in the composition of interest.
  • Human immune response refers to an immune response mediated by antibody molecules
  • cellular immune response is an immune response mediated by T-lymphocytes and/or other white blood cells.
  • the immune composition or vaccine of the present disclosure can achieve any one or a combination selected from the following: up-regulate the expression and activation of costimulatory molecules (eg CD80+, CD86+, CD40+) on dendritic cells
  • dendritic cells increase the expression of cytokines (such as TNF- ⁇ , IFN- ⁇ , KC, IFN- ⁇ ) in the lung, increase the titer of IgG antibodies (such as IgG1, IgG2a, IgG3), and increase IL-2 positive cells Proportion, increase the proportion of IFN- ⁇ positive cells, and reduce the viral load in the lungs.
  • costimulatory molecules eg CD80+, CD86+, CD40+
  • cytokines such as TNF- ⁇ , IFN- ⁇ , KC, IFN- ⁇
  • IgG antibodies such as IgG1, IgG2a, IgG3
  • IL-2 positive cells Proportion increase the proportion of IFN- ⁇ positive cells, and reduce the viral load in the
  • a method for preventing or treating coronavirus infection which comprises administering to a subject a preventive or therapeutically effective amount of the composition or vaccine of the present disclosure.
  • the administration of the composition is systemic (also systemic) or local.
  • the immune composition or vaccine of the present disclosure is administered by parenteral (eg, intramuscular, intraperitoneal, intravenous, subcutaneous, intradermal) injection.
  • parenteral eg, intramuscular, intraperitoneal, intravenous, subcutaneous, intradermal
  • the immune composition or vaccine of the present disclosure is delivered intracutaneously in a manner other than injection (for example, a manner that does not use a mechanical device to destroy the epithelial cell barrier).
  • the immune composition is administered by rectal, vaginal, nasal, oral, sublingual, respiratory, ocular, or transdermal routes.
  • the immune composition is administered intramuscularly.
  • the immune composition is administered intranasally.
  • the composition/vaccine of the present disclosure can be used in the form of an insufflator, nebulizer, pressure package, or introduction of a gas spray.
  • the pressure package can be provided with a suitable propellant (such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide).
  • a suitable propellant such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide.
  • the dosage unit can be determined by providing a gas valve.
  • the composition/vaccine of the present disclosure is a composition in the form of a dry powder, for example, a powdered mixture of a modifier and a suitable dry powder base such as lactose or starch.
  • the powder mixture can be packed in capsules and cartridges in the form of a unit dose, or, for example, in a gelatin package.
  • the powder can be administered from these containers by means of an inhaler or insufflator.
  • the antibody can be administered in liquid spray or nasal drops, such as a plastic sprayer.
  • the immune composition or vaccine of the present disclosure is administered to the subject at a frequency selected from: once a month, twice a month, 3 times a month, 4 times a month, 5 times a month , 6 times a month, 7 times a month, 8 times a month, 1 time a week, 2 times a week, 3 times a week, 4 times a week, 5 times a week, 6 times a week, 1 every three days Times, 2 times every three days, 3 times every three days, 1 time every two days, 2 times every two days, 1 time a day, 2 times a day.
  • the prevention or treatment method includes administering the immune composition of the present disclosure containing the SARS-CoV-2 antigen, and administering the composition of the nucleic acid encoding the SARS-CoV-2 antigen is also feasible.
  • Nucleic acid-based technology can directly apply naked or encapsulated nucleic acid encoding spike polypeptides to tissues and cells. This technology is based on the ability of these nucleic acids to be taken up and expressed by the cells of the recipient organism to generate immunogenic determinants.
  • this protein-based vaccine can induce a neutralizing and protective antibody-dependent immune response.
  • kits for implementing the above prevention or treatment method which comprises at least one container containing the immune composition or vaccine of the present disclosure.
  • the composition and/or amount of the composition in different containers can be the same or different.
  • the immune composition or vaccine of the present disclosure is formulated in a sterile liquid and packed in a sterile container (such as a tube, bottle, ampoule, syringe).
  • a sterile container such as a tube, bottle, ampoule, syringe.
  • the immune composition or vaccine of the present disclosure is contained in a container in the form of a dry powder or a freeze-dried powder. Before use, it is formulated into liquid form.
  • the kit of the present disclosure further comprises one or a combination selected from the following: needles, water for injection, instructions for use.
  • Figure 1 The pharmaceutical composition of the present disclosure activates the body's DC to up-regulate the production of costimulatory molecules.
  • Figures 2A to 2B can effectively increase the concentration of cytokines in the lungs after intranasal administration of the pharmaceutical composition of the present disclosure to mice.
  • Figure 3 IgG subtype antibody titers.
  • FIG. 4 Titer of IgG antibody against the new coronavirus S protein in rabbit serum.
  • Figure 5 The titers of neutralizing antibodies induced by different pharmaceutical compositions are different.
  • Figure 6A to Figure 6D Proportion of cells expressing IL-2 and IFN- ⁇ .
  • Figure 7 Viral load in the lungs of mice after challenge.
  • Figure 8 Viral load in the lungs of mice after challenge.
  • Antigen means any substance that can induce an immune response by being recognized by the host's immune system when it invades the host (for example, proteins, peptides, cancer cells, glycoproteins, glycolipids, live viruses, killed viruses, DNA etc.).
  • the antigen can be provided as a purified form or an unpurified form.
  • Antigens in this application include pathogen proteins, recombinant proteins, peptides, polysaccharides, glycoproteins, glycolipids and polynucleotides, live viruses, recombinant viruses, attenuated or inactivated viruses.
  • An immunogenic fragment refers to a part of the aforementioned "antigen" (for example, when the antigen is a protein or polypeptide, the immunogenic fragment is a non-full-length peptide fragment of the protein or polypeptide), which still retains (in whole or in part) the The host's immune system recognizes the ability to induce an immune response.
  • Inactivation refers to the removal of the pathogenicity and reproductive ability of the virus, but still retains the ability to stimulate the human body to produce an immune response.
  • the method of inactivating the virus is well known in the industry. Any common method can be used to inactivate the virus, and it can be appropriately selected according to the type of virus. These methods of inactivating viruses include, but are not limited to: the use of photoreactive compounds, oxidants, radiation (such as UV rays; ⁇ -rays), riboflavin and UV rays, and solvent-detergent treatment (such as organic solvents).
  • the attenuated virus is still alive, but the virus loses its ability to cause disease during the manufacturing process, but it still retains the ability to reproduce and stimulate the body to produce an immune response.
  • spike protein covers the category of its variants.
  • a variant of a spike protein refers to a polypeptide that is substantially homologous to the natural spike polypeptide, but has an amino acid sequence different from the natural spike protein due to the deletion, insertion or substitution of one or more amino acids.
  • Variants may contain conservatively substituted sequences.
  • a conservative substitution refers to the substitution of a given amino acid residue by a residue with similar biochemical properties. Such conservative substitutions are known.
  • the invention also includes naturally occurring spike polypeptide variants.
  • examples of such variants are proteins produced due to alternative mRNA splicing events or due to protease cleavage of spike polypeptides.
  • Variations attributable to proteolysis include, for example, differences in the ends that appear after expression in different types of host cells, which are caused by the proteolytic removal of one or more terminal amino acids from the spike polypeptide.
  • the SARS-CoV-2 antigen of the present disclosure is a protein or polypeptide
  • it can be prepared in the form of a multimer (homomultimer or heteromultimer), such as but not limited to dimers, trimers, Tetramers and pentamers, thereby providing more opportunities for antigen exposure.
  • the technician fuses a piece of foldable at the N or C-terminus of the SARS-CoV-2 antigen, and this piece of foldable helps the antigen to form a multimeric form.
  • Such a folder is well known in the art and does not affect the correct folding of the antigen in this application, nor does it affect the immunogenicity.
  • a foldon that can be used for trimerization when linked to a protein or peptide (also called a fusion), allows the formation of a trimer structure.
  • any known trimerization foldons/domains can be used. Examples of trimerization folds/domains include but are not limited to: HIV-1gp41 trimerization domain, SIV gp41 trimerization domain, Ebola virus gp-2 trimerization domain, HTLV-1gp-21 Trimerization domain, T4fibritin trimerization domain, yeast heat shock transcription factor trimerization domain, and human collagen trimerization domain.
  • CN111560074A can form twenty-tetramer nanoantigens through HP_Ferritin self-assembly; CN111607605A can self-assemble to form pentamers with the help of LTB26 to obtain pentamers of fusion proteins.
  • unit dose refers to a physically separate unit, suitable for use as a unit dose for an individual subject.
  • Each unit contains a predetermined amount of the composition of the present disclosure, and the predetermined amount presents an amount sufficient to produce the desired effect together with a pharmaceutically/physiologically acceptable diluent, carrier, or carrier.
  • the "effective amount" of the composition refers to the amount of the composition that is non-toxic but sufficient to provide the desired response (such as an immune response) and produce the corresponding preventive/therapeutic effect.
  • the exact amount required will vary depending on the subject It is different, depending on their species, age, the overall condition of the subject, the severity of the disease to be treated, and the specific macromolecule of interest, the method of administration, etc.
  • an appropriate "effective amount” can be determined by the present Those of ordinary skill in the art use conventional experimental methods to determine the effective amount for human subjects, for example, through animal studies.
  • treatment refers to obtaining the desired pharmacological and/or physiological effects.
  • the effect may be preventive from the perspective of completely and/or partially preventing the disease or its symptoms, and/or the effect may be caused by completely and/or partially stabilizing or curing the disease and/or disease
  • the negative effects can be of a medical nature.
  • compositions drugs, immune compositions, and vaccines can be used interchangeably.
  • Example 1 Preparation method of pharmaceutical composition 1 of the present disclosure
  • Antigen Recombinant new coronavirus S-trimer protein.
  • composition of the present disclosure according to the following composition:
  • Composition 1 of the present disclosure (unit dose 1ml) contains:
  • Recombinant new coronavirus S-trimer protein (2.5 ⁇ g, 5 ⁇ g, 6 ⁇ g, 10 ⁇ g and 20 ⁇ g), PIC (1mg/ml),
  • the above composition is formulated in a physiologically acceptable buffer (sodium chloride 150 mM, disodium hydrogen phosphate 0.0043 M and sodium dihydrogen phosphate 0.0017 M).
  • Example 2 Preparation method of pharmaceutical composition 2 of the present disclosure
  • Antigen Recombinant new coronavirus S-trimer protein.
  • composition of the present disclosure according to the following composition:
  • Composition 2 of the present disclosure (unit dose 1ml) contains:
  • Recombinant new coronavirus S-trimer protein (2.5 ⁇ g, 5 ⁇ g, 10 ⁇ g and 20 ⁇ g), PIC (1mg/ml),
  • Kanamycin 800 units/ml
  • Polysorbate 80 (0.01% w/v).
  • the above composition is formulated in a physiologically acceptable buffer (sodium chloride 150 mM, disodium hydrogen phosphate 0.0043 M and sodium dihydrogen phosphate 0.0017 M).
  • Antigen Recombinant new coronavirus S1 protein.
  • composition of the present disclosure according to the following composition:
  • Composition 3 of the present disclosure (unit dose 1ml) contains:
  • Recombinant new coronavirus S1 protein (2.5 ⁇ g, 5 ⁇ g, 6 ⁇ g, 10 ⁇ g and 20 ⁇ g), PIC (1mg/ml),
  • Kanamycin 800 units/ml
  • the above composition was formulated in a physiologically acceptable buffer (sodium chloride 150 mM, disodium hydrogen phosphate 0.0043 and sodium dihydrogen phosphate 0.0017 M).
  • Antigen Recombinant new coronavirus RBD protein (ie, the spike protein RBD region of SARS-CoV-2).
  • composition of the present disclosure according to the following composition:
  • Composition 4 of the present disclosure (unit dose 1ml) contains:
  • Recombinant new coronavirus RBD protein (2.5 ⁇ g, 5 ⁇ g, 6 ⁇ g, 10 ⁇ g and 20 ⁇ g), PIC (1mg/ml),
  • the above composition is formulated in a physiologically acceptable buffer (sodium chloride 150 mM, disodium hydrogen phosphate 0.0043 M and sodium dihydrogen phosphate 0.0017 M).
  • Example 5 Preparation method of pharmaceutical composition 5 of the present disclosure
  • Composition 5 of the present disclosure (unit dose 1ml) contains:
  • the above composition is formulated in a physiologically acceptable buffer (sodium chloride 150 mM, disodium hydrogen phosphate 0.0043 M and sodium dihydrogen phosphate 0.0017 M).
  • Antigen inactivated and purified new coronavirus.
  • composition 6 of the present disclosure (unit dose 1ml) according to the following composition, including:
  • Kanamycin 800 units/ml
  • Aluminum phosphate (aluminum content 0.5% w/w).
  • the above composition is formulated in a physiologically acceptable buffer (sodium chloride 150 mM, disodium hydrogen phosphate 0.0043 M and sodium dihydrogen phosphate 0.0017 M).
  • Test Example 1 The pharmaceutical composition 5 of the present disclosure activates dendritic cells to up-regulate costimulatory molecules
  • BMDC marrow-derived dendritic cells
  • results As shown in Figure 1, the ratio of CD80+ cells increased from 59.82% to 90.66%, the ratio of CD86+ cells increased from 11.70% to 38.66%, and the ratio of CD40+ cells increased from 16.98% after the action of the pharmaceutical composition 5 of the present disclosure with mouse BMDC. Increased to 58.62%, significantly up-regulating the production of costimulatory molecules.
  • Test Example 2 5 drops of the pharmaceutical composition of the present disclosure can effectively increase the concentration of cytokines in the lungs after nasal administration
  • mice were given PBS or the pharmaceutical composition 5 of the present disclosure via nasal drops, and were sacrificed at the specified time.
  • the lung tissues were collected, and the TNF- ⁇ , IFN- ⁇ , and KC of the samples were repeatedly tested with the Bio-plex protein array system; the IFN- ⁇ was detected with an ELISA kit.
  • mice After intranasal administration of the pharmaceutical composition 5 of the present disclosure to mice, the cytokines TNF- ⁇ , IFN- ⁇ , KC, and IFN- ⁇ in the lungs of the mice are significantly increased, indicating that the nasal administration of the mice can effectively activate the respiratory tract
  • the innate immunity causes immune cells to produce cytokines and chemokines. These cytokines and chemokines are related to the inhibition of mouse respiratory virus replication.
  • Test Example 3 The pharmaceutical composition 1 of the present disclosure can effectively induce the production of IgG antibodies (IgG1, IgG2a, IgG3) against the S1 protein of the new coronavirus in mice
  • mice were immunized with the pharmaceutical composition 1 of the present disclosure, blood was collected to separate the serum, and the titers of the IgG antibody subtypes against the new coronavirus S1 protein in the mouse serum were detected by the ELISA method.
  • the pharmaceutical composition of the present disclosure can significantly increase the IgG subtype antibody titer against the new coronavirus S1 protein in the serum ( Figure 3).
  • Test Example 4 The pharmaceutical composition 1 of the present disclosure can effectively induce rabbits to produce IgG antibodies against the new coronavirus S protein
  • the rabbits were immunized with the pharmaceutical composition 1 of the present disclosure, the blood was collected to separate the serum, and the IgG antibody titer against the new coronavirus S protein in the rabbit's serum was detected by the ELISA method.
  • the pharmaceutical composition of the present disclosure can significantly increase the titer of IgG antibody against the new coronavirus S protein in rabbit serum ( Figure 4).
  • Test Example 5 The pharmaceutical compositions 1, 3-4 of the present disclosure can effectively induce rabbits to produce neutralizing antibodies against neocoronavirus
  • Rabbits were immunized with pharmaceutical compositions 1, 3-4 of the present disclosure, blood was collected to separate serum, and the neutralizing antibody titer of rabbit serum was detected by pseudovirus neutralization test.
  • compositions 1, 3-4 of the present disclosure can significantly increase the titers of neutralizing antibodies in rabbit serum, and the titers of neutralizing antibodies induced by different pharmaceutical compositions are different (Figure 5).
  • Test Example 6 The pharmaceutical composition 2 of the present disclosure effectively induces a new coronavirus-specific cellular immune response in mice
  • mice were immunized with the pharmaceutical composition 2 of the present disclosure, the spleen was taken to separate the spleen cell suspension, and the Elispot method was used to detect the antigen-specific cellular immune response.
  • composition 2 of the present disclosure can significantly improve the cellular immune response of mice, and significantly increase the proportion of cells expressing IL-2 and IFN- ⁇ (see Figure 6A to Figure 6D for the results).
  • Test Example 7 The pharmaceutical composition 1 of the present disclosure effectively reduces the new coronavirus load in hACE2 mice
  • Method Use the pharmaceutical composition 1 of the present disclosure to immunize hACE2 transgenic mice (6 weeks old, 18-26g, SARS-CoV-2 infected hACE2 transgenic mouse model), and use SARS-CoV-2 14 days after the initial immunization A nasal drip challenge was performed, and lung tissue was taken 5 days after the challenge to test the lung viral load.
  • the pharmaceutical composition 1 of the present disclosure can significantly reduce the viral load in the lungs of mice after challenge, relieve lung inflammation to a certain extent, and has a good preventive effect (see Figure 7 for the results).
  • Test Example 8 The pharmaceutical composition 2 of the present disclosure effectively reduces the new coronavirus load in hACE2 mice
  • SARS-CoV-2 was used to challenge hACE2 mice by nasal drip, and the pharmaceutical composition 2 of the present disclosure was administered at the same time, and lung tissues were taken 5 days after the challenge to detect the viral load of the lungs.
  • composition 2 of the present disclosure can significantly reduce the viral load in the lungs of mice after challenge, and has a good therapeutic effect (see Figure 8 for the results).
  • Test Example 9 The pharmaceutical composition 2 of the present disclosure can effectively induce the production of IgG antibodies against the new coronavirus S protein in cynomolgus monkeys
  • the cynomolgus monkeys were immunized with the pharmaceutical composition 2 of the present disclosure, blood was collected to separate the serum, and the IgG antibody titer against the new coronavirus S protein in the cynomolgus monkey serum was detected by the ELISA method.
  • the pharmaceutical composition of the present disclosure can significantly increase the IgG antibody titer against the new coronavirus S protein in the serum of cynomolgus monkeys.
  • Animals in the low-dose pharmaceutical composition 2 of the present disclosure and the high-dose pharmaceutical composition 2 of the present disclosure can detect specific IgG antibodies one week after the second immunization (D15, before the third dose), with an incidence of 10 /10 and 10/10, the antibody titers ranged from 1:1600 to >1:102400.
  • the titers of animal-specific IgG antibodies in the low-dose and high-dose groups increased.
  • the titers of all animal-specific IgG antibodies in the low-dose and high-dose groups did not decrease.
  • Test Example 10 The pharmaceutical composition 6 of the present disclosure can effectively induce rabbits to produce neutralizing antibodies against the new coronavirus
  • the rabbits were immunized with the pharmaceutical composition 6 of the present disclosure, blood was collected to separate the serum, and the neutralizing antibody titers in the rabbit serum were detected by the SARS-CoV-2 field strain neutralization test.
  • composition 6 of the present disclosure can induce neutralizing antibody titers in rabbit serum.
  • the drug combination for preventing and treating SARS-CoV-2 infection of the novel coronavirus provided by the present disclosure can achieve the following effects:
  • the components of the drug combination of the present disclosure can activate interferon regulatory factor 3 and NF- ⁇ B, up-regulate the expression of type I interferon and pro-inflammatory factors, activate the body's natural immunity and acquired immunity, and play an important role The antiviral effect.
  • the results of multiple animal experiments show that the components of the pharmaceutical composition of the present disclosure can effectively reduce the titer load of animal respiratory viruses.
  • the dosage form is flexible, and the injection dosage form can be used.
  • the effect is fast and reliable.
  • the liquid medicine is directly injected into the tissues, and is not affected by pH, enzymes, food, etc., has no first-pass effect, and the drug content is not easy to lose. Therefore, the curative effect is reliable and can be used to rescue patients in crisis.
  • Spray preparations can also be used, which are convenient for medication, easy for patients to accept, rapid transmucosal absorption, quick onset of action, and good patient compliance.

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Abstract

本申请涉及包含聚核苷酸的药物组合物及其用于预防或治疗COVID-19的用途。更具体地，本申请公开了一种用于预防或治疗COVID-19的组合物，其包含聚核糖肌苷-聚核糖胞苷酸、抗生素或多氨基化合物、正价离子以及任选地源自新型冠状病毒SARS-CoV-2的抗原，还涉及上述组合物在制备预防或治疗新型冠状病毒SARS-CoV-2感染的药物或疫苗中的用途。

Description

包含聚核苷酸的药物组合物及其用于预防或治疗COVID-19的用途
本申请要求2020年05月29日提交的中国专利申请“包含聚核苷酸的药物组合物及其用于预防或治疗COVID-19的用途”(申请号202010475388.6)的优先权。
技术领域
本申请涉及医学领域，尤其是冠状病毒感染的预防或治疗。更具体而言，涉及一种用于预防或治疗冠状病毒感染的药物组合物，其含有：聚核糖肌苷-聚核糖胞苷酸(PIC)、抗生素(或多氨基化合物)、正价离子、以及任选地源自病毒的抗原(例如但不限于：病毒颗粒、蛋白、核酸、肽)；以及该组合物用于预防或治疗冠状病毒感染的用途。
背景技术
新型冠状病毒感染主要是新型冠状病毒(SARS-CoV-2，也作2019-nCOV)所致。传染途径主要为经呼吸道飞沫传播，亦可通过接触传播。人群普遍易感，老年人及有基础疾病者感染后病情较重，儿童及婴幼儿也有发病。
基于目前的流行病学调查，COVID-19潜伏期1-14天，多为3-7天。以发热、干咳、乏力为主要表现。少数患者伴有鼻塞、流涕、咽痛、肌痛和腹泻等症状。重症患者多在发病一周后出现呼吸困难和/或低氧血症，严重者可快速进展为急性呼吸窘迫综合征、脓毒症休克、难以纠正的代谢性酸中毒和出凝血功能障碍及多器官衰竭等。值得注意的是重症、危重症患者病程中可为中低热，甚至无明显发热。轻型患者仅表现为低热、轻微乏力等，无肺炎表现。从目前病例情况看，多数患者预后良好，少数患者病情危重。老年人和有慢性基础疾病者预后较差。儿童病例症状相对较轻。目前所见传染源主要是新型冠状病毒感染的患者。无症状感染者也可能成为传染源。病毒传播能力强，没有有效药物，危害极大。
针对COVID-19的治疗，目前没有确认有效的抗病毒治疗方法。大多数候选药物仅限于体外实验，还未在动物或人体中证实其治疗效果。因而，本领域仍需要提供一种有效的抗病毒治疗组合物。
发明内容
免疫组合物
鉴于此，本公开提供了一种免疫组合物，其包含聚核糖肌苷-聚核糖胞苷酸(polyI:C，PIC)、抗生素(或多氨基化合物)、正价离子以及任选源自病毒的抗原；或者，由PIC、抗生素(或多氨基化合物)、正价离子以及任选病毒抗原组成。
在一些实施方式中，所述的病毒是冠状病毒，例如SARS-CoV-2(涵盖其变种或亚种)。在一些实施方式中，所述的病毒是灭活粗制的、或者是灭活纯化型的。
在一些实施方式中，适用于本公开的疫苗或免疫组合物，包括但不限于灭活型、亚单位型、基因重组型、多肽型疫苗、核酸型。
在一些实施方式中，适用于本申请的PIC可以是市售PIC，或者是本领域(例如2006800007767和2006800010596)中公开的基于聚肌苷酸-聚胞苷酸的佐剂，其通过引用全部并入此处。
在一些实施方式中，免疫组合物中PIC在分子量上是异质的，其分子量为至少50,000道尔顿。例如，66,000道尔顿的数值相当于6.4S沉降系数单位(Svedbergs)的分子尺寸。在一些实施方式中，PIC为66,000道尔顿至1,200,000道尔顿(相当于6.4至24.0沉降系数单位)。在其它一些实施方式中，PIC的分子量是至少150,000道尔顿。在其它一些实施方式中，PIC的分子量是100,000至200,000道尔顿，或是300,000至4,000,000道尔顿，或是500,000至1,000,000道尔顿，或是1,000,000至1,500,000道尔顿，或是1,500,000至2,000,000道尔顿，或是2,000,000至2,500,000道尔顿，或是2,500,000至3,000,000道尔顿，或是3,000,000至3,500,000道尔顿，或是3,500,000至4,000,000道尔顿，或是4,000,000至4,500,000道尔顿，或是4,500,000至5,000,000道尔顿。
本公开中所用术语“异质”，是指免疫组合物或疫苗中所含的PIC分 子，在分子量、尺寸或二者皆有的物理特性上不是均一的。
在一些实施方式中，所述PIC在免疫组合物/疫苗中浓度为250μg/单位剂量至5000μg/单位剂量；例如，所述PIC在组合物中浓度选自：250μg/单位剂量、500μg/单位剂量、1000μg/单位剂量、1500μg/单位剂量、2000μg/单位剂量、3000μg/单位剂量、4000μg/单位剂量、5000μg/单位剂量，以及上述任意两点之间的范围。
在具体的实施方式中，PIC在免疫组合物/疫苗中浓度为500μg/单位剂量至4000μg/单位剂量，1000μg/单位剂量至3000μg/单位剂量，或者1000μg/单位剂量至2500μg/单位剂量。
当本公开的免疫组合物/疫苗适用于人时，PIC在免疫组合物/疫苗中浓度选自500μg/单位剂量、1000μg/单位剂量、1500μg/单位剂量、2000μg/单位剂量以及上述任意两点之间的范围。当本公开的免疫组合物/疫苗适用于未成年人(如儿童)时，PIC在免疫组合物/疫苗中浓度选自250μg/单位剂量、500μg/单位剂量、1000μg/单位剂量、1250μg/单位剂量以及上述任意两点之间的范围。
在具体的实施方式中，所含的抗生素选自：泰百霉素、蒽环霉素、丁苷菌素硫酸盐、庆大霉素、潮霉素、艾米康丝菌素、卡那霉素(或双去氧卡那霉素)、暗霉素、美它酰胺、新霉素、嘌呤霉素、链霉素、链脲霉素及其组合。所述的多氨基化合物选自：亚精氨酸盐、精眯、N-(3-氨基丙基)、N-(3-氨基丙基)-1,4-丁二胺、精碱精素、精素、OS-二甲基胺基硫代磷酸酯、多聚赖氨酸、氨基糖苷及其组合。在一些实施方案中，所述抗生素在组合物中的浓度为10单位/ml至100,000单位/ml，优选100单位/ml至10,000单位/ml，更优选500单位/ml至5,000单位/ml。
在一些实施方案中，抗生素或多氨基化合物在免疫组合物中的浓度可以提及的是400U/单位剂量至1200U/单位剂量，例如但不限于400、500、600、700、800、900、1000、1100、1200U/单位剂量。
在另一些实施方案中，抗生素或多氨基化合物在免疫组合物中的浓度可以提及的是400U/ml至1200U/ml，例如但不限于400、500、600、700、800、900、1000、1100、1200U/ml。
在一些实施方式中，所含的正价离子为二价正价离子，并且选自： 钙、镉、锂、镁、铈、铯、铬、钴、氘、镓、碘、铁、锌及其组合。在具体的实施方式中，所述的正价离子为钙离子。所述正价离子可以呈任何适当的盐或有机复合物的形式，包括但不限于：氯化物、氟化物、氢氧化物、磷酸盐或硫酸盐。例如，当该正价离子是钙时，则钙离子可呈碳酸钙、氯化钙、氟化钙、氢氧化钙、磷酸钙或硫酸钙的形式。在一些实施方案中，所述的正价离子在组合物中的浓度为10μmol至10mmol/ml，例如50μmol至5mmol/ml；或者100μmol至1mmol/ml。
在一些实施方案中，正价离子在免疫组合物中的浓度为0.01mg/单位剂量至0.1mg/单位剂量，例如但不限于0.01、0.02、0.03、0.04、0.05、0.06、0.07、0.08、0.09、0.1mg/单位剂量。
在另一些实施方案中，正价离子在免疫组合物中的浓度为0.01mg/ml至0.1mg/ml，例如但不限于0.01、0.02、0.03、0.04、0.05、0.06、0.07、0.08、0.09、0.1mg/ml。
在一些实施方式中，源自冠状病毒的抗原是病毒(例如，减毒、灭活病毒、人工组装的病毒载体或病毒颗粒)。
在一些具体的实施方式中，源自冠状病毒的抗原是灭活病毒SARS-CoV-2、减毒的病毒SARS-CoV-2、或在受试者中不能繁殖的病毒SARS-CoV-2。所述病毒相对于PIC的比例选自：1活性单位/50μg、1活性单位/60μg、1活性单位/70μg、1活性单位/80μg、1活性单位/90μg、1活性单位/100μg、1活性单位/125μg、1活性单位/200μg、1活性单位/250μg、1活性单位/300μg、1活性单位/350μg、1活性单位/400μg、1活性单位/450μg、1活性单位/500μg、1活性单位/550μg、1活性单位/600μg、1活性单位/700μg、1活性单位/800μg、1活性单位/1000μg、1活性单位/1500μg、1活性单位/2000μg、1活性单位/2500μg、1活性单位/3000μg、1活性单位/4000μg、1活性单位/5000μg、1活性单位6000μg、1活性单位/7000μg、1活性单位/8000μg、1活性单位/9000μg、1活性单位/10000μg以及上述任意两点之间的范围。
在一些实施方式中，当源自冠状病毒的抗原是病毒(例如，减毒、灭活病毒、人工组装的病毒载体或病毒颗粒)时，所述抗原在组合物中的浓度为0.2活性单位/单位剂量至100活性单位/单位剂量。更具体地，所述病毒或源自病毒的抗原在组合物中的浓度选自0.2活性单位/ 单位剂量、0.5活性单位/单位剂量、1.0活性单位/单位剂量、1.5活性单位/单位剂量、2.0活性单位/单位剂量、2.5活性单位/单位剂量、3.0活性单位/单位剂量、3.5活性单位/单位剂量、4.0活性单位/单位剂量、5.0活性单位/单位剂量、6.0活性单位/单位剂量、7.0活性单位/单位剂量、8.0活性单位/单位剂量、9.0活性单位/单位剂量、10.0活性单位/单位剂量、15.0活性单位/单位剂量、20.0活性单位/单位剂量、30.0活性单位/单位剂量、40.0活性单位/单位剂量、50.0活性单位/单位剂量、60.0活性单位/单位剂量、70.0活性单位/单位剂量、80.0活性单位/单位剂量、90.0活性单位/单位剂量、100.0活性单位/单位剂量，以及上述任意两点之间的范围。在具体的实施方式中，所述病毒或源自病毒的抗原在组合物中的浓度为0.5活性单位/单位剂量至3.0活性单位/单位剂量；优选1.0活性单位/单位剂量至2.5活性单位/单位剂量。
在一些实施方式中，当源自冠状病毒的抗原是病毒(例如，减毒、灭活病毒、人工组装的病毒载体或病毒颗粒)时，所述抗原在组合物中的浓度为0.05活性单位/ml至40.0活性单位/ml；优选0.05活性单位/ml、0.1活性单位/ml、0.15活性单位/ml、0.2活性单位/ml、0.5活性单位/ml、1.0活性单位/ml、2.0活性单位/ml、3.0活性单位/ml、4.0活性单位/ml、5.0活性单位/ml、10活性单位/ml、15活性单位/ml、20活性单位/ml、25活性单位/ml、30活性单位/ml、35活性单位/ml、40活性单位/ml。
在一些实施方式中，当所述源自冠状病毒的抗原选自以下时：SARS-CoV-2的S蛋白或其免疫原性片段、M蛋白或其免疫原性片段、N蛋白或其免疫原性片段、E蛋白或其免疫原性片段、依据SARS-CoV-2结构设计的蛋白、多肽、DNA、RNA，所述抗原相对于所述PIC的比例选自：1μg/10μg、1μg/20μg、1μg/30μg、1μg/40μg、1μg/50μg、1μg/60μg、1μg/70μg、1μg/80μg、1μg/90μg、1μg/100μg、1μg/125μg、1μg/200μg、1μg/250μg、1μg/300μg、1μg/350μg、1μg/400μg、1μg/450μg、1μg/500μg、1μg/550μg、1μg/600μg、1μg/700μg、1μg/800μg、1μg/1000μg、1μg/1500μg、1μg/2000μg、1μg/2500μg、1μg/3000μg、1μg/4000μg、1μg/5000μg、1μg/6000μg、1μg/7000μg、1μg/8000μg、1μg/9000μg、1μg/10000μg以及上述任意两点之间的范围。
在一些实施方案中，当所述源自冠状病毒的抗原选自以下时：SARS-CoV-2的S蛋白或其免疫原性片段、M蛋白或其免疫原性片段、N蛋白或其免疫原性片段、E蛋白或其免疫原性片段、依据SARS-CoV-2结构设计的蛋白、多肽、DNA、RNA，其在免疫组合物中的浓度为0.1μg/单位剂量至1000.0μg/单位剂量，例如但不限于0.5μg/单位剂量、1.0μg/单位剂量、2.0μg/单位剂量、3.0μg/单位剂量、4.0μg/单位剂量、5.0μg/单位剂量、6.0μg/单位剂量、7.0μg/单位剂量、8.0μg/单位剂量、9.0μg/单位剂量、10.0μg/单位剂量、15.0μg/单位剂量、20.0μg/单位剂量、30.0μg/单位剂量、40.0μg/单位剂量、50.0μg/单位剂量、60.0μg/单位剂量、70.0μg/单位剂量、80.0μg/单位剂量、90.0μg/单位剂量、100.0μg/单位剂量、200.0μg/单位剂量、300.0μg/单位剂量、400.0μg/单位剂量、500.0μg/单位剂量、600.0μg/单位剂量、700.0μg/单位剂量、800.0μg/单位剂量、900.0μg/单位剂量、1000.0μg/单位剂量，以及上述任意两点之间的范围；或者例如0.1μg/ml至1000.0μg/ml；优选1μg/ml、2μg/ml、3μg/ml、5μg/ml、10μg/ml、20μg/ml、30μg/ml、40μg/ml、50μg/ml、100μg/ml、200μg/ml、500μg/ml、1000μg/ml。
在一些实施方式中，源自冠状病毒的抗原是SARS-CoV-2刺突蛋白时，抗原在免疫组合物中的浓度是2.5μg/单位剂量至20μg/单位剂量。
在一些实施方式中，源自冠状病毒的抗原是SARS-CoV-2刺突蛋白多聚体时(例如同源三聚体)，抗原在免疫组合物中的浓度是2.5μg/单位剂量至20μg/单位剂量。
在一些实施方式中，源自冠状病毒的抗原是SARS-CoV-2刺突蛋白的免疫原性片段时，抗原在免疫组合物中的浓度是2.5μg/单位剂量至20μg/单位剂量。
在一些实施方式中，源自冠状病毒的抗原是SARS-CoV-2刺突蛋白RBD区时，抗原在免疫组合物中的浓度是2.5μg/单位剂量至20μg/单位剂量。
在一些实施方式中，本公开中所述的单位剂量以体积的形式表示，并且选自：0.1ml、0.15ml、0.2ml、0.5ml、1.0ml、1.5ml、2.0ml、2.5ml、3.0ml、4.0ml、5.0ml、10.0ml、20.0ml、30.0ml、40.0ml、50.0ml、60.0ml、70.0ml、80.0ml、90.0ml、100.0ml，150ml，200ml， 250.00ml，以及上述任意两点之间的范围。技术人员理解，过大或过小的单位剂量导致临床操作不便。因此，当本公开的免疫组合物/疫苗注射施用于人类受试者时，单位剂量优选在0.5ml至1.0ml范围内。当本公开的免疫组合物/疫苗鼻内施用于人类受试者时，单位剂量优选在0.15ml至0.2ml范围内。当本公开的免疫组合物/疫苗静脉注射于人类受试者时，单位剂量优选在30.0ml至1000ml范围内。此处应当理解的是，单位剂量虽然以体积进行表示，但是这并不意味着本公开的免疫组合物/疫苗只能是液体形式。当本公开的免疫组合物/疫苗制备成固体(干粉或冻干粉)时，单位剂量的体积是指干粉或冻干粉重新配制后的体积。
在一些实施方案中，本公开的免疫组合物或疫苗可以包含明胶、蔗糖、白砂糖、乳糖、麦芽糖、海藻糖、葡萄糖、低分子右旋糖苷、山梨醇、聚山梨醇酯20、聚山梨醇酯80、盐酸精氨酸、甘露醇聚乙二醇、人血白蛋白、重组白蛋白、辛酸钠、尿素、氢氧化铝、酚红、磷酸铝、角鲨烯、皂苷、寡核苷酸、氯化镁、氯化钾、氯化钠、硫代硫酸钠、磷酸二氢钾、抗坏血酸、三氯甲烷、苯酚、硫柳汞。
在一些实施方式中，本公开的免疫组合物或疫苗还可以包含生理上可接受的缓冲液，选自：乙酸盐、Tris、碳酸氢盐、碳酸盐、磷酸盐缓冲液。适用于本公开组合物的缓冲液pH值选自：6.50、6.60、6.70、6.80、6.90、7.00、7.05、7.1、7.15、7.2、7.25、7.30、7.35、7.40、7.45、7.50、7.55、7.60、7.65、7.70、7.75、7.80、7.85、7.90、7.95、8.00以及上述任意两点之间的范围。在具体的实施方式中，缓冲液是PBS，并且pH值在7.0至8.0范围之内。
在一些实施方式中，缓冲液在组合物中浓度是5mM至50mM，具体地5mM至20mM。可以提及的示例是5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32、33、34、35、36、37、38、39、40、41、42、43、44、45、46、47、48、49、50mM。
缓冲液在制备成本公开的组合物或疫苗后，不排除其pH会随着其他成分的加入而发生变化。鉴于缓冲液的缓冲能力，在一些实施方式中，本公开的免疫组合物或疫苗的pH值被控制在适当的范围，例如： 6.0、6.1、6.2、6.3、6.4、6.5、6.6、6.7、6.8、6.9、7.0、7.05、7.1、7.15、7.2、7.25、7.3、7.35、7.4、7.45、7.5、7.55、7.6、7.65、7.7、7.75、7.8、7.85、7.9、7.95、8.0、8.1、8.2、8.3、8.4、8.5、8.6、8.7、8.8、8.9、9.0以及上述任意两点之间的范围。在具体的实施方式中，组合物的pH值在7.0至8.0范围之内。
本公开的免疫组合物或疫苗可被制备成为干燥粉末、液态溶液(例如可注射型溶液、水性或生理食盐水溶液，或悬浮液、油膏、液滴、乳剂、凝胶、糖浆或浆液、气雾剂、喷雾剂、滴鼻剂)、片剂、包衣片剂、微胶囊、栓剂、药片、颗粒、糖衣锭、胶囊。制备方法被一般性地叙述在Stanley A Plotkin等人，Vaccine第四版W.B.Saunders Company 2003中。优选，本公开的免疫组合物或疫苗被制备成为可注射型溶液。
根据本公开的另一方面，提供一种免疫组合物，其是上述免疫组合物所对应的干粉或冻干粉形式。技术人员能够理解的是，当免疫组合物是干粉或冻干粉形式时，和液体形式的免疫组合物区别仅在于含水量。作为一个示例，当干粉或冻干粉形式的免疫组合物包含2mg/ml PIC时，技术人员知晓这个浓度是指将免疫组合物重新配制成液体形式后PIC的浓度。
在一些实施方案中，提供一种免疫组合物，其包含：
源自冠状病毒的抗原、
0.5mg/ml至2mg/ml PIC、
400单位/ml至1200单位/ml卡那霉素、
0.01mg/ml至0.1mg/ml氯化钙、
100mM至200mM氯化钠、
5mM至20mM磷酸缓冲液。
在一些实施方案中，提供一种免疫组合物，其包含：
源自冠状病毒的抗原、
0.5mg/ml至2mg/ml PIC、
400单位/ml至1200单位/ml卡那霉素、
0.01mg/ml至0.1mg/ml氯化钙、
100mM至200mM氯化钠、
5mM至20mM磷酸缓冲液、
任选地，100mM至200mM盐酸精氨酸、
任选地，0.005％w/v至0.05％w/v聚山梨醇酯80、
任选地，0.1％w/w至1.0％w/w磷酸铝。
方法和用途
根据本公开的另一方面，提供了上述PIC、抗生素(或多氨基化合物)以及正价离子的组合在制备预防或治疗冠状病毒感染的药物或疫苗中的用途。
根据本公开的又一方面，提供了CN103405762A中所公开的聚核苷酸佐剂组合物在制备预防或治疗冠状病毒感染的药物或疫苗中的用途。
根据本公开的另一方面，提供了PIC、抗生素(或多氨基化合物)、正价离子和病毒的组合在制备预防或治疗冠状病毒感染的药物或疫苗中的用途。
根据本公开的另一方面，提供了本公开的免疫组合物或疫苗在预防或治疗冠状病毒感染中的用途。
更具体地，提供了本公开的免疫组合物或疫苗在制备预防或治疗冠状病毒感染的药物中的用途。
根据本公开的另一方面，还涉及在宿主动物中刺激免疫反应的方法，该方法包括向该动物给予有效诱导免疫反应的量的本文所述免疫组合物。所述宿主动物较佳是哺乳动物，更佳是猴，仍更佳是人。本发明还涉及使宿主动物针对病毒产生免疫的方法，该方法包括向所述动物给予其量能有效诱导保护性反应的本文所述免疫组合物。所述宿主动物较佳是哺乳动物，更佳是猴，还更佳是人。
对抗原或组合物的“免疫反应”是在对象的体液和/或细胞免疫反应中产生的针对存在于感兴趣的组合物中的分子的反应。“体液免疫反应”指抗体分子介导的免疫反应，而“细胞免疫反应”是由T-淋巴细胞和/或其它白细胞介导的免疫反应。
在一些具体的实施方案中，本公开的免疫组合物或疫苗能够实现选自以下的任一项或组合：上调共刺激分子(例如CD80+、CD86+、CD40+)在树突状细胞上的表达、激活树突状细胞、提高细胞因子(例如TNF-α、IFN-γ、KC、IFN-β)在肺中的表达、提高IgG抗体(例如 IgG1、IgG2a、IgG3)滴度、增加IL-2阳性细胞比例、增加IFN-γ阳性细胞比例、降低肺部病毒载量。
根据本公开的另一方面，提供了一种预防或治疗冠状病毒感染的方法，其包括向受试者施用预防或治疗有效量的本公开组合物或疫苗。
在一些实施方式中，组合物的施用是系统性的(也作全身性)或局部的。
在一些实施方式中，通过胃肠道外(如肌肉内、腹腔内、静脉内、皮下、皮内)注射来施用本公开的免疫组合物或疫苗。
在其它实施方式中，本公开的免疫组合物或疫苗是以注射以外的方式(例如不以机械装置来破坏上皮细胞障壁的方式)经皮内传输。
在另一些实施方式中，免疫组合物是通过直肠、阴道、鼻、口、舌下、呼吸道、眼、或透皮的途径而施用。
在一些实施方式中，免疫组合物是通过肌肉内施用。
在一些实施方式中，免疫组合物是滴鼻施用。
在一些实施方式中，对于吸入给药，本公开的组合物/疫苗可以用吹入器、雾化器、压力包装或导入气体喷雾的方式。压力包装可以带有合适的推进剂(如二氯二氟甲烷、三氯一氟甲烷、二氯四氟乙烷、二氧化碳)。在用加压气雾剂的情况下，可以通过提供气阀来确定剂量单位。
可选性地，对于吸入或吹入给药，本公开的组合物/疫苗是干粉形式的组合物，例如，调节剂和合适的干粉基质如乳糖或淀粉的粉状混合物。粉状混合物可以以单位剂量的形式装于胶囊和药筒中，或例如明胶包装中，药粉可以借助吸入器或吹入器从这些容器中给药。对于鼻腔给药，抗体可以通过液体喷雾或滴鼻剂型，例如通过一个塑料喷雾器。
在一些实施方式中，按照选自以下的频率向受试者施用本公开的免疫组合物或疫苗：每月1次、每月2次、每月3次、每月4次、每月5次、每月6次、每月7次、每月8次、每周1次、每周2次、每周3次、每周4次、每周5次、每周6次、每三天1次、每三天2次、每三天3次、每两天1次、每两天2次、每天1次、每天2次。
在一些实施方式中，预防或治疗方法包括施用包含SARS-CoV-2 抗原的本公开的免疫组合物，施用编码SARS-CoV-2抗原的核酸的组合物也可行。本领域技术人员熟知核酸疫苗和核酸疫苗技术以及基于蛋白质和多肽的技术的概念、应用、以及有效性。基于核酸的技术能够将裸露的或者包囊化的编码刺突多肽的核酸直接施用至组织和细胞。该技术是基于这些核酸能够被受者生物体的细胞摄取并表达以产生免疫原性决定簇。在单次或者多次注射本公开的免疫组合物(例如含SARS-CoV-2的刺突蛋白)，这种基于蛋白质的疫苗可诱导中和性、保护性抗体依赖性免疫应答。
根据本公开的另一方面，提供了一种用于实施上述预防或治疗方法的药盒，其包含至少一个容器，所述容器包含本公开的免疫组合物或疫苗。位于不同容器中组合物的组成和/或量可以相同，也可以不同。
在一些实施方式中，本公开的免疫组合物或疫苗配制于无菌液体内，并且装在无菌容器(如管、瓶、安瓿、注射器)内。在另一些实施方式中，本公开的免疫组合物或疫苗以干粉或冻干粉的形式装在容器内。临用前，配制成液体形式。
在一些实施方式中，本公开的药盒还包含选自以下的一项或其组合：针头、注射用水、使用说明书。
附图说明
图1本公开药物组合物激活机体DC上调共刺激分子产生。
图2A至图2B滴鼻给予小鼠本公开药物组合物后能够有效提高肺内部细胞因子的浓度。
图3：IgG亚型抗体滴度。
图4：家兔血清中抗新冠病毒S蛋白的IgG抗体滴度。
图5：不同的药物组合物所诱导产生的中和抗体滴度不同。
图6A至图6D：表达IL-2、IFN-γ的细胞比例。
图7：小鼠攻毒后，肺部病毒载量。
图8：小鼠攻毒后，肺部病毒载量。
具体实施方式
“抗原”意指当其入侵宿主时，能够通过被宿主的免疫系统识别 而诱导免疫应答的任何物质(例如，蛋白、肽、癌症细胞、糖蛋白、糖脂、活病毒、杀死的病毒、DNA等)。抗原可以作为纯化的形式或未纯化的形式提供。本申请中的抗原，包括病原体的蛋白、重组蛋白、肽、多糖、糖蛋白、糖脂和多聚核苷酸、活病毒、重组的病毒、减毒的或灭活的病毒。
免疫原性片段是指前述“抗原”的一部分(例如，当抗原是蛋白或多肽时，免疫原性片段是该蛋白或多肽的非全长肽片段)，其仍保留了(全部或部分)被宿主的免疫系统识别而诱导免疫应答的能力。
灭活，是指去除病毒的致病能力和繁殖能力，但是仍然保留了刺激人体产生免疫应答的能力。使病毒灭活的方法为业界所熟知。任何常用方法均可使用以使病毒灭活，且可针对病毒种类而适当地选用。这些使病毒灭活的方法包括但不限于：利用光反应性化合物、氧化剂、辐射(例如UV射线；γ-射线)、核黄素和UV射线的组合、溶剂-去污剂处理(例如以有机溶剂三-N-丁基-磷酸酯和Tween 80去污剂来处理)、聚乙二醇处理、巴斯德灭菌法(热处理)、低pH处理、酶处理(胃蛋白酶或胰蛋白酶)、亚甲蓝光处理、以二甲基亚甲蓝和可见光进行处理、以补骨脂内酯衍生物S-59和UVA照射进行处理。
减毒，减毒的病毒仍然是活的，但是在制造过程中使病毒丧失致病能力，但是仍然保留繁殖能力和刺激人体产生免疫应答的能力。
本文中，术语刺突蛋白(Spike或S蛋白)涵盖其变体范畴。刺突蛋白的变体是指与天然刺突多肽实质上同源的多肽，但由于一个或多个氨基酸的缺失、插入或取代，而具有不同于天然刺突蛋白的氨基酸序列。变体可包含保守性取代的序列，保守性取代是指一种给定的氨基酸残基被具有相似生化特性的残基取代。此类保守性置换是已知的，保守性取代的实例包括脂肪族残基彼此间的置换，例如Ile、Val、Leu、或Ala彼此之间的取代；或者是极性残基彼此之间的置换，例如Lys与Arg之间；Glu与Asp之间；和Gln与Asn之间。本发明也包括天然存在的刺突多肽变体。此类变体的实例是因替代性mRNA剪接事件或因刺突多肽的蛋白酶裂解而产生的蛋白质。可归因于蛋白水解作用的变异包括例如在不同类型的宿主细胞内表达后出现的末端的差异，这是由于自刺突多肽蛋白水解去除了一或多个末端氨基酸所致。
当本公开的SARS-CoV-2抗原是蛋白或多肽时，可以制备成多聚体的形式(同源多聚体或异源多聚体)，例如但不限于二聚体、三聚体、四聚体、五聚体，从而提供更多的抗原接触机会。在具体的示例中，技术人员在SARS-CoV-2抗原的N或C端融合一段折叠子，这段折叠子有助于抗原形成多聚体的形式。这样的折叠子在本领域中是公知的且不影响抗原本申请的正确折叠，也不影响免疫原性。作为示例，可用于三聚化的折叠子，当与蛋白质或肽连接(也称为融合)时允许形成三聚体结构。在本公开中，可以使用任何已知的三聚化折叠子/结构域。三聚化折叠子/结构域的例子包括但不限于：HIV-1gp41三聚化结构域、SIV gp41三聚化结构域、埃博拉病毒gp-2三聚化结构域、HTLV-1gp-21三聚化结构域、T4fibritin三聚化结构域、酵母热休克转录因子三聚化结构域、和人胶原三聚化结构域。再比如，CN111560074A中可通过HP_Ferritin自组装作用形成二十四聚体纳米抗原；CN111607605A借助LTB26可自组装形成五聚体的特性，得到融合蛋白的五聚体。
本公开中所用术语“单位剂量”，是指在实体上呈分离的单位，适合作为使用于受试者个体的单位剂量。各个单位含有预定量的本公开组合物，该预定量呈现足以和药学/生理上可接受的稀释剂、载剂或载体共同产生所需效应的用量。
本文术语，组合物的“有效量””是指无毒但足以提供所需反应(如免疫反应)和产生相应的预防/治疗效果的组合物的量。所需的确切量随受试者不同而不同，依据它们的物种、年龄、对象的总体条件、要治疗的病症的严重程度、以及感兴趣的具体大分子、给药的方式等。在具体的示例中合适的“有效量”可以由本领域的普通技术人员使用常规实验方法进行测定，例如通过动物研究确定人受试者的有效量。
本说明书中所用术语“治疗”是指，获得所需药理和/或生理效应。该效应从完全地和/或部分地防止疾病或其症状的角度来看可以是属于预防性质的，和/或该效应从完全地和/或部分地稳定或治愈疾病和/或疾病所导致的负面效果来看可以是属于医疗性质的。
本说明书中，当描述数值范围时，所用的表述“…至…”、“范围之内”或“范围之间”包含端点值。
本说明书的上下文中，药物组合物、药物、免疫组合物、疫苗可 互换使用。
实施例
如无特别说明，本公开的试验动物操作遵照《实验动物管理条例》、《关于善待实验动物的指导性意见》、国家标准GB/14925进行。
实施例1：本公开药物组合物1的制备方法
1.抗原：重组新冠病毒S-三聚体蛋白。
2.按照以下组成制备本公开的组合物：
本公开的组合物1(单位剂量1ml)包含：
重组新冠病毒S-三聚体蛋白(2.5μg、5μg、6μg、10μg和20μg)、PIC(1mg/ml)、
卡那霉素(800单位/ml)、和
氯化钙(0.0472mg/ml)。
在无菌条件下，将上述组合物配制在生理可接受的缓冲液(氯化钠150mM、磷酸氢二钠0.0043M和磷酸二氢钠0.0017M)中。
表1
Figure PCTCN2021096048-appb-000001
实施例2：本公开药物组合物2的制备方法
1.抗原：重组新冠病毒S-三聚体蛋白。
2.按照以下组成制备本公开的组合物：
本公开的组合物2(单位剂量1ml)包含：
重组新冠病毒S-三聚体蛋白(2.5μg、5μg、10μg和20μg)、PIC(1mg/ml)、
卡那霉素(800单位/ml)、
氯化钙(0.0472mg/ml)、
盐酸精氨酸(140mM)、
聚山梨醇酯80(0.01％w/v)。
在无菌条件下，将上述组合物配制在生理可接受的缓冲液(氯化钠150mM、磷酸氢二钠0.0043M和磷酸二氢钠0.0017M)中。
表2
Figure PCTCN2021096048-appb-000002
实施例3：本公开药物组合物3的制备方法
1.抗原：重组新冠病毒S1蛋白。
2.按照以下组成制备本公开的组合物：
本公开的组合物3(单位剂量1ml)包含：
重组新冠病毒S1蛋白(2.5μg、5μg、6μg、10μg和20μg)、PIC(1mg/ml)、
卡那霉素(800单位/ml)、
氯化钙(0.0472mg/ml)。
在无菌条件下，将上述组合物配制在生理可接受的缓冲液(氯化 钠150mM、磷酸氢二钠0.0043和磷酸二氢钠0.0017M)中。
表3
Figure PCTCN2021096048-appb-000003
实施例4：本公开药物组合物4的制备方法
1.抗原：重组新冠病毒RBD蛋白(即，SARS-CoV-2的刺突蛋白RBD区)。
2.按照以下组成制备本公开的组合物：
本公开的组合物4(单位剂量1ml)包含：
重组新冠病毒RBD蛋白(2.5μg、5μg、6μg、10μg和20μg)、PIC(1mg/ml)、
卡那霉素(800单位/ml)、和
氯化钙(0.0472mg/ml)。
在无菌条件下，将上述组合物配制在生理可接受的缓冲液(氯化钠150mM、磷酸氢二钠0.0043M和磷酸二氢钠0.0017M)中。
表4
Figure PCTCN2021096048-appb-000004
Figure PCTCN2021096048-appb-000005
实施例5：本公开药物组合物5的制备方法
本公开的组合物5(单位剂量1ml)包含：
PIC(1mg/ml)、
卡那霉素(800单位/ml)和
氯化钙(0.0472mg/ml)。
在无菌条件下，将上述组合物配制在生理可接受的缓冲液(氯化钠150mM、磷酸氢二钠0.0043M和磷酸二氢钠0.0017M)中。
表5
Figure PCTCN2021096048-appb-000006
实施例6：本公开药物组合物6的制备方法
1.抗原：灭活纯化新冠病毒。
2.按照以下组成制备本公开的组合物6(单位剂量1ml)，包含：
灭活纯化新冠病毒(600活性单位)、
PIC(1mg/ml)、
卡那霉素(800单位/ml)、
氯化钙(0.0472mg/ml)、和
磷酸铝(铝含量0.5％w/w)。
在无菌条件下，将上述组合物配制在生理可接受的缓冲液(氯化钠150mM、磷酸氢二钠0.0043M和磷酸二氢钠0.0017M)中。
表6
Figure PCTCN2021096048-appb-000007
测试例1：本公开药物组合物5激活机体树突状细胞上调共刺激分子
方法：通过采用C57BL/6小鼠髓源树突状细胞(BMDC)体外与本公开药物组合物5共同孵育18小时。流式细胞仪检测CD80+、CD86+和CD40+细胞的比例和平均荧光强度。
结果：如图1所示本公开药物组合物5与小鼠BMDC作用后CD80+细胞的比例从59.82％提升至90.66％，CD86+细胞的比例从11.70％提升至38.66％，CD40+细胞的比例从16.98％提升至58.62％，显著地上调共刺激分子产生。
结论：通过本实验证明，本公开药物组合物5能够显著诱导机体树突状细胞上调共刺激分子，促进树突状细胞的活化。
测试例2：本公开药物组合物5滴鼻给药后能够有效提高肺内部细胞因子的浓度
方法：小鼠分别通过滴鼻给予PBS或本公开药物组合物5，并在指定时间处死。采集肺部组织，采用Bio-plex蛋白阵列系统对样品的TNF-α、IFN-γ及KC进行重复测试；采用ELISA试剂盒检测IFN-β。
表7.实验动物分组
Figure PCTCN2021096048-appb-000008
结果：见图2A至图2B。
结论：滴鼻给予小鼠本公开药物组合物5后，小鼠肺部的细胞因子TNF-α、IFN-γ、KC、IFN-β显著升高，表明经鼻给药小鼠可有效激活呼吸道的固有免疫，导致免疫细胞产生细胞因子和趋化因子，这些细胞因子和趋化因子与抑制小鼠呼吸道病毒复制有关。
测试例3：本公开药物组合物1可以有效诱导小鼠产生抗新冠病毒S1蛋白的IgG抗体(IgG1、IgG2a、IgG3)产生
方法：分别采用本公开药物组合物1对小鼠进行免疫，取血分离血清，采用ELISA法检测小鼠血清中抗新冠病毒S1蛋白的IgG抗体亚型的滴度。
表8.实验动物分组
Figure PCTCN2021096048-appb-000009
结论：本公开药物组合物可以显著提高血清中抗新冠病毒S1蛋白的IgG亚型抗体滴度(图3)。
测试例4：本公开药物组合物1可以有效诱导家兔产生抗新冠病毒S蛋白的IgG抗体产生
分别采用本公开药物组合物1对家兔进行免疫，取血分离血清，采用ELISA法检测家兔血清中抗新冠病毒S蛋白的IgG抗体滴度。
表9.实验动物分组
Figure PCTCN2021096048-appb-000010
结论：本公开药物组合物可以显著提高家兔血清中抗新冠病毒S蛋白的IgG抗体滴度(图4)。
测试例5：本公开药物组合物1、3-4可以有效诱导家兔产生抗新 冠病毒中和抗体产生
方法：分别采用本公开药物组合物1、3-4对家兔进行免，取血分离血清，采用假病毒中和试验检测家兔血清中中和抗体滴度。
表10.实验动物分组
Figure PCTCN2021096048-appb-000011
结论：本公开药物组合物1、3-4可显著提高家兔血清中的中和抗体滴度，且不同的药物组合物所诱导产生的中和抗体滴度不同(图5)。
测试例6：本公开药物组合物2有效诱导小鼠产生新冠病毒特异性细胞免疫反应产生
方法：分别采用本公开药物组合物2对小鼠进行免疫，取脾分离脾细胞悬液，采用Elispot法检测抗原特应性细胞免疫反应。
表11.实验动物分组
Figure PCTCN2021096048-appb-000012
结论：本公开药物组合物2可以显著提高小鼠细胞免疫反应，显著增加表达IL-2、IFN-γ的细胞比例(结果见图6A至图6D)。
测试例7：本公开药物组合物1有效降低hACE2小鼠中的新冠病毒载量
方法：采用本公开药物组合物1对hACE2转基因小鼠(6周龄，18-26g，SARS-CoV-2感染hACE2转基因小鼠模型)进行免疫，于初 免后14天采用SARS-CoV-2进行滴鼻攻毒，于攻毒后5天取肺组织检测肺部病毒载量。
表12.实验动物分组
Figure PCTCN2021096048-appb-000013
结论：本公开药物组合物1可以显著降低小鼠攻毒后肺部病毒载量，一定程度缓解肺部炎症，具有良好的预防效果(结果见图7)。
测试例8：本公开药物组合物2有效降低hACE2小鼠中的新冠病毒载量
方法：采用SARS-CoV-2对hACE2小鼠进行滴鼻攻毒，同时给予本公开药物组合物2，于攻毒后5天取肺组织检测肺部病毒载量。
表13.实验动物分组
Figure PCTCN2021096048-appb-000014
结论：本公开药物组合物2可以显著降低小鼠攻毒后肺部病毒载量，具有良好的治疗效果(结果见图8)。
测试例9：本公开药物组合物2可以有效诱导食蟹猴产生抗新冠病毒S蛋白的IgG抗体产生
方法：分别采用本公开药物组合物2对食蟹猴进行免疫，取血分离血清，采用ELISA法检测食蟹猴血清中抗新冠病毒S蛋白的IgG抗体滴度。
表14.实验动物分组
Figure PCTCN2021096048-appb-000015
结论：本公开药物组合物可以显著提高食蟹猴血清中抗新冠病毒S蛋白的IgG抗体滴度。本公开药物组合物2低剂量和本公开药物组合物2高剂量组动物在第二次免疫后1周(D15，第3次给药前)可检测出特异性IgG抗体，发生率分别为10/10和10/10，抗体滴度范围均为1:1600至>1:102400。随给药次数的增加，低剂量和高剂量组动物特异性IgG抗体滴度均可见增加。停药恢复4周后(D57)，低剂量和高剂量组所有动物特异性IgG抗体滴度均未见降低。
测试例10：本公开药物组合物6可以有效诱导家兔产生抗新冠病毒中和抗体产生
方法：分别采用本公开药物组合物6对家兔进行免疫，取血分离血清，采用SARS-CoV-2野毒株中和试验检测家兔血清中的中和抗体滴度。
表15.实验动物分组
Figure PCTCN2021096048-appb-000016
结论：本公开药物组合物6可诱导家兔血清中的中和抗体滴度。
通过本公开提供的预防和治疗新型冠状病毒SARS-CoV-2感染的药物组合，能够达到如下效果：
1.药效好，疗效显著，本公开药物组合成分可激活干扰素调节因 子3及NF-κB，上调I型干扰素及前炎症因子的表达，激活人体的天然免疫和获得性免疫，发挥重要的抗病毒作用。多个动物试验结果显示本公开药物组合成分能够有效降低动物呼吸道病毒滴度载量。
2.可同时起到预防和治疗效果。
3.剂型灵活，可采用注射剂型，作用迅速可靠其药液直接注入组织，不受pH、酶、食物等影响，无首过效应，药物含量不易损失，因此疗效可靠，可用于抢救危机病人。亦可采用喷雾制剂，用药方便，患者易于接受，经粘膜吸收迅速、起效快、患者依从性好。

Claims (14)

  1. 一种免疫组合物，其包含：
    聚核糖肌苷-聚核糖胞苷酸(PIC)、
    抗生素或多氨基化合物、
    正价离子、以及
    任选地，源自冠状病毒的抗原；
    所述冠状病毒是SARS-CoV-2或其变种；
    优选地，所述抗生素选自以下的一项或组合：泰百霉素、蒽环霉素、丁苷菌素硫酸盐、庆大霉素、潮霉素、艾米康丝菌素、卡那霉素、暗霉素、β-内酰胺、美它酰胺、新霉素、嘌呤霉素、链霉素、链脲霉素；
    优选地，所述多氨基化合物选自以下的一项或组合：亚精氨酸盐、精眯、N-(3-氨基丙基)、N-(3-氨基丙基)-1,4-丁二胺、精碱精素、精素、OS-二甲基胺基硫代磷酸酯、多聚赖氨酸、氨基糖苷；
    优选地，所述正价离子为二价正价离子，更优选地，所述正价离子选自以下的一项或组合：钙、镉、锂、镁、铈、铯、铬、钴、氘、镓、碘、铁、锌；最优选，所述正价离子为钙离子；
    优选地，所述源自冠状病毒的抗原选自以下的一项或组合：灭活病毒SARS-CoV-2、减毒的病毒SARS-CoV-2、在受试者中不能繁殖的病毒SARS-CoV-2、SARS-CoV-2的S蛋白或其免疫原性片段、M蛋白或其免疫原性片段、N蛋白或其免疫原性片段、E蛋白或其免疫原性片段、依据SARS-CoV-2结构设计的蛋白、多肽、RNA、DNA。
  2. 根据权利要求1所述的免疫组合物，其中：
    当所述源自冠状病毒的抗原是灭活病毒SARS-CoV-2、减毒的病毒SARS-CoV-2、或在受试者中不能繁殖的病毒SARS-CoV-2时，所述源自冠状病毒的抗原相对于所述PIC的比例选自以下的任一项：
    1活性单位/50μg、1活性单位/60μg、1活性单位/70μg、1活性单位/80μg、1活性单位/90μg、1活性单位/100μg、1活性单位/125μg、1活性单位/200μg、1活性单位/250μg、1活性单位/300μg、1活性单位/350μg、1活性单位/400μg、1活性单位/450μg、1活性单位/500μg、1 活性单位/550μg、1活性单位/600μg、1活性单位/700μg、1活性单位/800μg、1活性单位/1000μg、1活性单位/1500μg、1活性单位/2000μg、1活性单位/2500μg、1活性单位/3000μg、1活性单位/4000μg、1活性单位/5000μg、1活性单位6000μg、1活性单位/7000μg、1活性单位/8000μg、1活性单位/9000μg、1活性单位/10000μg以及上述任意两点之间的范围；
    或者，当所述源自冠状病毒的抗原选自以下的任一项时：SARS-CoV-2的S蛋白或其免疫原性片段、M蛋白或其免疫原性片段、N蛋白或其免疫原性片段、E蛋白或其免疫原性片段、依据SARS-CoV-2结构设计的蛋白、多肽、DNA、RNA，所述抗原相对于所述PIC的比例选自以下的任一项：
    1μg/10μg、1μg/20μg、1μg/30μg、1μg/40μg、1μg/50μg、1μg/60μg、1μg/70μg、1μg/80μg、1μg/90μg、1μg/100μg、1μg/125μg、1μg/200μg、1μg/250μg、1μg/300μg、1μg/350μg、1μg/400μg、1μg/450μg、1μg/500μg、1μg/550μg、1μg/600μg、1μg/700μg、1μg/800μg、1μg/1000μg、1μg/1500μg、1μg/2000μg、1μg/2500μg、1μg/3000μg、1μg/4000μg、1μg/5000μg、1μg/6000μg、1μg/7000μg、1μg/8000μg、1μg/9000μg、1μg/10000μg以及上述任意两点之间的范围。
  3. 根据权利要求1所述的免疫组合物，其中：
    所述PIC在免疫组合物中的浓度为250μg/单位剂量至5000μg/单位剂量；
    优选，所述PIC在免疫组合物中的浓度选自以下的任一项：250μg/单位剂量、500μg/单位剂量、1000μg/单位剂量、1500μg/单位剂量、2000μg/单位剂量、3000μg/单位剂量、4000μg/单位剂量、5000μg/单位剂量。
  4. 根据权利要求1所述的免疫组合物，其中：
    当所述源自冠状病毒的抗原是灭活病毒SARS-CoV-2、减毒的病毒SARS-CoV-2、或在受试者中不能繁殖的病毒SARS-CoV-2时，所述源自冠状病毒的抗原在免疫组合物中的浓度为0.1活性单位/单位剂量至 100.0活性单位/单位剂量；
    优选地，选自以下的任一项：0.5活性单位/单位剂量、1.0活性单位/单位剂量、1.5活性单位/单位剂量、2.0活性单位/单位剂量、2.5活性单位/单位剂量、3.0活性单位/单位剂量、3.5活性单位/单位剂量、4.0活性单位/单位剂量、5.0活性单位/单位剂量、6.0活性单位/单位剂量、7.0活性单位/单位剂量、8.0活性单位/单位剂量、9.0活性单位/单位剂量、10.0活性单位/单位剂量、15.0活性单位/单位剂量、20.0活性单位/单位剂量、30.0活性单位/单位剂量、40.0活性单位/单位剂量、50.0活性单位/单位剂量、60.0活性单位/单位剂量、70.0活性单位/单位剂量、80.0活性单位/单位剂量、90.0活性单位/单位剂量、100.0活性单位/单位剂量，以及上述任意两点之间的范围；
    或者，当所述源自冠状病毒的抗原选自以下的任一项时：SARS-CoV-2的S蛋白或其免疫原性片段、M蛋白或其免疫原性片段、N蛋白或其免疫原性片段、E蛋白或其免疫原性片段、依据SARS-CoV-2结构设计的蛋白、多肽、DNA、RNA，其在免疫组合物中的浓度为0.1μg/单位剂量至1000.0μg/单位剂量；
    优选地，选自以下的任一项：0.5μg/单位剂量、1.0μg/单位剂量、2.0μg/单位剂量、3.0μg/单位剂量、4.0μg/单位剂量、5.0μg/单位剂量、6.0μg/单位剂量、7.0μg/单位剂量、8.0μg/单位剂量、9.0μg/单位剂量、10.0μg/单位剂量、15.0μg/单位剂量、20.0μg/单位剂量、30.0μg/单位剂量、40.0μg/单位剂量、50.0μg/单位剂量、60.0μg/单位剂量、70.0μg/单位剂量、80.0μg/单位剂量、90.0μg/单位剂量、100.0μg/单位剂量、200.0μg/单位剂量、300.0μg/单位剂量、400.0μg/单位剂量、500.0μg/单位剂量、600.0μg/单位剂量、700.0μg/单位剂量、800.0μg/单位剂量、900.0μg/单位剂量、1000.0μg/单位剂量，以及上述任意两点之间的范围。
  5. 根据权利要求3或4所述的免疫组合物，其中
    所述单位剂量选自以下的任一项：0.1ml，0.15ml，0.2ml，0.5ml，1.0ml，1.5ml，2.0ml，2.5ml，3.0ml，4.0ml，5.0ml，10.0ml，20.0ml，30.0ml，40.0ml，50.0ml，60.0ml，70.0ml，80.0ml，90.0ml， 100.0ml，150.0ml，200.0ml，250.0ml，以及上述任意两点之间的范围。
  6. 根据权利要求1至5中任一项所述的免疫组合物，其还包含选自以下的一种或其组合：
    明胶、蔗糖、白砂糖、乳糖、麦芽糖、海藻糖、葡萄糖、低分子右旋糖苷、山梨醇、聚山梨醇酯20、聚山梨醇酯80、盐酸精氨酸、甘露醇聚乙二醇、人血白蛋白、重组白蛋白、辛酸钠、尿素、氢氧化铝、磷酸铝、角鲨烯、皂苷、寡核苷酸、酚红、氯化镁、氯化钾、氯化钠、硫代硫酸钠、磷酸二氢钾、抗坏血酸、三氯甲烷、苯酚、硫柳汞。
  7. 根据权利要求1至6中任一项所述的免疫组合物，其还包含生理上可接受的缓冲液，选自以下的一种或其组合：乙酸盐、三羟甲基氨基甲烷、碳酸氢盐、碳酸盐、磷酸盐缓冲液；
    优选地，缓冲液浓度是5mM至50mM，优选5mM至20mM；
    优选地，所述免疫组合物的pH值为6至9。
  8. 根据权利要求1至7中任一项所述的免疫组合物，可被制备成为固体剂型或液态剂型；
    所述液态剂型选自以下的任一项：注射溶液、悬浮液、喷雾剂、气雾剂、滴鼻剂、油膏、乳剂、滴剂、糖浆剂、凝胶剂；
    所述固体剂型选自以下的任一项：干粉、冻干粉、片剂、胶囊剂、栓剂、颗粒剂、糖衣锭。
  9. 根据权利要求1至8中任一项所述的免疫组合物，其中：
    所述PIC在分子量上是异质的，其分子量为50,000道尔顿至1,200,000道尔顿；或者
    所述PIC在分子量上是异质的，其沉降系数单位为5.0至24.0。
  10. 根据权利要求1至9中任一项所述的免疫组合物，其中：
    所述抗生素或多氨基化合物在免疫组合物中的浓度为400U/单位剂量至1200U/单位剂量，优选400U/单位剂量、500U/单位剂量、600 U/单位剂量、700U/单位剂量、800U/单位剂量、900U/单位剂量、1000U/单位剂量、1100U/单位剂量、1200U/单位剂量；
    所述正价离子在免疫组合物中的浓度为0.01mg/单位剂量至0.1mg/单位剂量，优选0.01mg/单位剂量、0.02mg/单位剂量、0.03mg/单位剂量、0.04mg/单位剂量、0.05mg/单位剂量、0.06mg/单位剂量、0.07mg/单位剂量、0.08mg/单位剂量、0.09mg/单位剂量、0.1mg/单位剂量。
  11. 根据权利要求1至10中任一项所述的免疫组合物，其中所述源自冠状病毒的抗原选自以下的任一项：
    2.5μg/单位剂量至20μg/单位剂量SARS-CoV-2刺突蛋白、
    2.5μg/单位剂量至20μg/单位剂量SARS-CoV-2刺突蛋白多聚体、
    2.5μg/单位剂量至20μg/单位剂量SARS-CoV-2刺突蛋白的免疫原性片段、
    2.5μg/单位剂量至20μg/单位剂量SARS-CoV-2刺突蛋白RBD区、
    300活性单位/单位剂量至1000活性单位/单位剂量灭活的SARS-CoV-2病毒体；
    优选地，所述多聚体是二聚体、三聚体或四聚体；
    优选地，所述多聚体是同源多聚体。
  12. 根据权利要求1至11中任一项所述的免疫组合物，其包含：
    源自冠状病毒的抗原、
    0.5mg/ml至2mg/ml PIC、
    400单位/ml至1200单位/ml卡那霉素、
    0.01mg/ml至0.1mg/ml氯化钙、
    100mM至200mM氯化钠、
    5mM至20mM磷酸缓冲液、
    任选地，还包含选自以下的任一项或组合：100mM至200mM盐酸精氨酸、0.005％w/v至0.05％w/v聚山梨醇酯80、0.1％w/w至1.0％w/w磷酸铝。
  13. 权利要求1至12中任一项所述的免疫组合物在制备用于预防 或治疗冠状病毒感染的疫苗或药物中的用途；
    优选地，所述冠状病毒是SARS-CoV-2或其变种；
    优选地，所述疫苗或药物制备成适于选自以下任一施用途径的剂型：肌肉内、腹腔内、静脉内、皮下、透皮、皮内、口服、舌下、呼吸道(鼻、咽、气管、支气管)。
  14. 一种预防或治疗冠状病毒SARS-CoV-2感染的方法，其包括：
    向受试者施用预防或治疗有效量的权利要求1至12中任一项所述的免疫组合物；
    优选地，按照以下的频率向所述受试者施用所述免疫组合物：
    每4年1至4次、每3年1至3次、每2年1至2次、每年1次、每年2次、每年3次、每年4次、每年5次、每年6次、每月1次、每月2次、每月3次、每月4次、每月5次、每月6次、每月7次、每月8次、每周1次、每周2次、每周3次、每周4次、每周5次、每周6次、每三天1次、每三天2次、每三天3次、每两天1次、每两天2次、每天1次、每天2次；
    每次施用之间的时间间隔是相同的或不同的；
    所述施用是选自以下任一项的施用途径：肌肉内、腹腔内、静脉内、皮下、透皮、皮内、口服、舌下、呼吸道(鼻、咽、气管、支气管)。
PCT/CN2021/096048 2020-05-29 2021-05-26 包含聚核苷酸的药物组合物及其用于预防或治疗covid-19的用途 Ceased WO2021238982A1 (zh)

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US17/768,441 US20230081039A1 (en) 2020-05-29 2021-05-26 Pharmaceutical composition comprising polynucleotides and use thereof for prevention or treatment of covid-19
CU2022000068A CU20220068A7 (es) 2020-05-29 2021-05-26 Composición inmune que contiene ácido polirriboinosínico-polirribocitidílico, un antibiótico o un compuesto poliamino, y un ion positivo
CA3178162A CA3178162A1 (en) 2020-05-29 2021-05-26 Pharmaceutical composition comprising polynucleotides and use thereof for prevention or treatment of covid-19
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EP21812811.4A EP4159233A4 (en) 2020-05-29 2021-05-26 POLYNUCLEOTIDE PHARMACEUTICAL COMPOSITION AND USE THEREOF FOR PREVENTING OR TREATING COVID-19
CN202180004682.1A CN114340662A (zh) 2020-05-29 2021-05-26 包含聚核苷酸的药物组合物及其用于预防或治疗covid-19的用途
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