CN113425832A

CN113425832A - Use of interferon lambda in the treatment of infections with novel coronaviruses (2019-nCoV)

Info

Publication number: CN113425832A
Application number: CN202010204993.XA
Authority: CN
Inventors: 杨柳; 李岩; 冯静; 郭万军; 潘海
Original assignee: Sciwind Biosciences Co Ltd
Current assignee: Sciwind Biosciences Co Ltd
Priority date: 2020-03-23
Filing date: 2020-03-23
Publication date: 2021-09-24

Abstract

The invention relates to application of IFN-lambda in preparing a medicine for treating patients infected by new coronavirus, which can effectively reduce the level of the new coronavirus in infected patients, and has more remarkable performance in controlling cytokine storm caused in the treatment process and relieving respiratory distress syndrome caused by the cytokine storm.

Description

Use of interferon lambda in the treatment of infections with novel coronaviruses (2019-nCoV)

The technical field is as follows:

the present invention relates to the use of interferon lambda in the treatment of novel coronavirus (2019-nCoV) infections.

Background art:

the novel coronavirus pneumonia is an acute respiratory infectious disease which is brought into a B infectious disease specified in infectious disease prevention and treatment Law of the people's republic of China, and is managed according to the A infectious disease.

The WHO formally named the novel coronavirus pneumonia as COVID-19(corona virus disease 2019). The international committee for virus classification named this virus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Coronaviruses are a class of enveloped single-stranded positive-stranded RNA viruses that infect a wide variety of mammals, including humans. Clinically, the virus is found to cause lung diseases firstly and further cause a plurality of systemic diseases. High frequency recombination and high mutation rates are considered to be the main reasons for the easy adaptation of coronaviruses to new hosts and niches. Prior to the outbreak of COVID-19, 6 types of coronaviruses are known to infect humans and cause respiratory disease. Among them, coronavirus (SARS-CoV) and the respiratory syndrome coronavirus (MERS-CoV) of middle east of 2012 are lethal viruses, and HCoV-OC43, HCoV-229E, HCoV-NL63 and HCoV-HKU1 coronaviruses cause only mild upper respiratory symptoms.

The coronaviridae family consists of 4 genera of α, β, γ, and δ. SARS-CoV-2 is a coronavirus of the beta genus, which is found by sequencing the virus isolated from the lower respiratory tract of patients with pneumonia. Coronaviruses are mainly composed of spike surface glycoprotein (S), small envelope protein (E), matrix protein (M), and nucleocapsid protein (N). Wherein the S protein can be divided into two functional subunits (S1 and S2), the S1 subunit promotes viral infection by binding to host receptors, and the S2 subunit promotes membrane fusion; the N protein plays a key role in viral transcription and assembly. It was found that the homology of SARS-CoV-2 with the gene sequence of SARS-CoV-like gene is about 80%, while the homology with two bat-derived coronaviruses (bat-SL-CoVZC45 and bat-SL-CoVZXC21) is as high as 88%. The related data indicate that SARS-CoV-2 is highly similar to the gene sequence of bat SARS-like coronavirus, presumably the natural host of SARS-CoV-2 is likely to be bat. The homology of SARS-CoV-2 and coronavirus separated from pangolin scales is found to reach 99% in the laboratory of agricultural university P3 in China Huazhong, suggesting that pangolin scales may be intermediate hosts.

Early studies have shown that an increase in proinflammatory cytokines such as Interleukin (IL) -1 β, IL-6, IL-12, interferon γ (IFN- γ), etc., in serum is associated with the development of inflammation and extensive lung injury [20 ]. Clinically, the levels of inflammatory factors such as IL-7, IL-8, IL-10, IFN-gamma, TNF-alpha and the like in the plasma of SARS-CoV-2 infected patients are higher than those of healthy adults, and ICU patients are higher than non-ICU patients, indicating that the cytokine storm is related to the severity of the disease.

Supportive treatment of patients is generally a standard treatment regimen, as no effective antiviral treatment has been established. When the medicine is taken, corresponding medicines are analyzed and selected mainly according to the clinical condition of a patient. Drugs that have been reported for patients with infections include antiviral drugs such as ritonavir (remdesivir), lopinavir/ritonavir (lopinavir-ritonavir), antifungal drugs, hormonal drugs, traditional Chinese medicine formulations, and symptomatic support measures.

The novel coronavirus pneumonia diagnosis and treatment scheme also provides that the alpha-interferon can be tried to be atomized and inhaled. Since interferon alpha has a wide distribution of its receptors in the body, influenza-like symptoms such as fever and fatigue are likely to occur in clinical hepatitis c and b treatments, and therefore, these side effects are likely to aggravate the symptoms of the novel coronavirus pneumonia infection in clinical treatments. And risks triggering a cytokine storm.

Human innate immunity plays a very important role as the first line of defense against viral infection. Whereas interferons play a more important role in innate immunity against viral infections. The interferons found in the early days are mainly type I (subtypes include α, β and ω) and type II (mainly γ) interferons, and a new interferon family found in 2003 as type III interferons is called interferon λ, and includes 3 subtypes of λ 1(IL29), λ 2(IL28A) and λ 3(IL28B), and λ 4 found later, and these subtypes have high sequence homology. With the further intensive research on the type III interferon family in recent years, it has been found that interferon λ plays a more important role in resisting viral infection of the body, and that its receptor distribution specificity has better efficacy and safety in antiviral therapy.

Interferon lambda receptors are mainly distributed in respiratory and intestinal epithelial cells, liver, B cells of immune cells, neutrophils, macrophages, plasmacytoid dendritic cells, and the like. However, bone marrow-derived hematopoietic stem cells (e.g., NK cells) and the like do not have receptor distribution, and therefore, side effects such as hematological and nervous system suppression and the like due to interferon alpha treatment are not involved.

Summary of The Invention

The object of the present invention is to provide a therapeutic method for treating a novel coronavirus (201-nCoV) infection with interferon lambda. More and more medical researches find that in the treatment process of novel coronavirus pneumonia, antiviral treatment and reduction of inflammation caused by cytokine storm are very critical, and the cytokine storm caused in the antiviral treatment process causes respiratory distress syndrome (ARDS).

The inventors of the present invention have surprisingly found that interferon lambda can simultaneously achieve inhibition of viral replication and inhibition of inflammatory damage caused by neutrophils. Has unique advantages compared with the drugs for resisting the new coronavirus which are already applied to clinic.

Accordingly, an aspect of the present invention provides the use of interferon λ for the manufacture of a medicament for the treatment of a novel coronavirus (2019-nCoV).

In a second aspect, the present invention provides a pharmaceutical composition, which comprises interferon lambda and a pharmaceutically acceptable excipient or adjuvant.

In the present invention, the interferon lambda (or referred to as interferon lambda polypeptide, IFN lambda polypeptide) is meant to include lambda 1(IL29), lambda 2(IL28A) and lambda 3(IL28B) or lambda 4, or a variant of said lambda 1, lambda 2, lambda 3 or lambda 4, which may be a deletion, insertion or substitution of one or more amino acids thereof, but still retains the biological activity of said lambda 1, lambda 2, lambda 3 or lambda 4 interferon.

The methods of the invention can comprise administering at least one IFN λ polypeptide, or a mixture of IFN- λ 1 and IFN- λ 2, or a mixture of IFN- λ 1 and IFN- λ 3, or a mixture of IFN- λ 2 and IFN- λ 3, or a mixture of IFN- λ 1, IFN- λ 2, and IFN- λ 3 can be administered to a patient.

By "FN lambda polypeptide" we include those polypeptides IFN-. lambda.1 (SEQ ID NO: 1), IFN-. lambda.2 (SEQ ID NO: 2), IFN-. lambda.3 (SEQ ID NO: 3) disclosed in GenBank accession Nos. Q8IU54, Q8IZJ0, Q8IZI9, and further IFN-. lambda.4 has a sequence as set forth in SEQ ID NO: 4, respectively.

IFN-λ1：

MAAAWTVVLVTLVLGLAVAGPVPTSKPTTTGKGCHIGRFKSLSPQELASFKKARDALE ESLKLKNWSCSSPVFPGNWDLRLLQVRERPVALEAELALTLKVLEAAAGPALEDVLDQPLH TLHHILSQLQACIQPQPTAGPRPRGRLHHWLHRLQEAPKKESAGCLEASVTFNLFRLLTRDL KYVADGNLCLRTSTHPEST

IFN-λ2：

MKLDMTGDCTPVLVLMAAVLTVTGAVPVARLHGALPDARGCHIAQFKSLSPQELQAF KRAKDALEESLLLKDCRCHSRLFPRTWDLRQLQVRERPMALEAELALTLKVLEATADTDPA LVDVLDQPLHTLHHILSQFRACIQPQPTAGPRTRGRLHHWLYRLQEAPKKESPGCLEASVTF NLFRLLTRDLNCVASGDLCV

IFN-λ3：

MTGDCMPVLVLMAAVLTVTGAVPVARLRGALPDARGCHIAQFKSLSPQELQAFKRAK DALEESLLLKDCKCRSRLFPRTWDLRQLQVRERPVALEAELALTLKVLEATADTDPALGDV LDQPLHTLHHILSQLRACIQPQPTAGPRTRGRLHHWLHRLQEAPKKESPGCLEASVTFNLFR LLTRDLNCVASGDLCV

IFN-λ4：

MRPSVWAAVAAGLWVLCTVIAAAPRRCLLSHYRSLEPRTLAAAKALRDRYEEEALSW GQRNCSFRPRRDPPRPSSCARLRHVARGIADAQAVLSGLHRSELLPGAGPILELLAAAGRDV AACLELARPGSSRKVPGAQKRRHKPRRADSPRCRKASVVFNLLRLLTWELRLAAHSGPCL

By administering an effective amount of an interferon lambda polypeptide to a patient infected with a novel coronavirus, the level of virus in the patient infected with the novel coronavirus is reduced, damage to airway epithelial cells caused by neutrophils is reduced, and respiratory distress syndrome caused during antiviral therapy is alleviated.

Interferon lambda is combined with a suitable pulmonary delivery vehicle for the treatment of novel coronavirus (201-nCoV) infections. Suitable carriers may be buffer systems, preservatives, surface tension agents, chelating agents, stabilizers and surfactants. In some embodiments, the medicaments or formulations of the present invention are aqueous medicaments or formulations, for example, they may be in the form of solutions or suspensions. In a particular embodiment of the invention, the drug or formulation is a stable aqueous solution. In other embodiments of the invention, the medicament or formulation is a lyophilized formulation to which a solvent and/or diluent is added prior to use.

In one embodiment of the invention, a drug delivery device is designed which comprises an inhaler of a liquid (e.g. a suspension or droplets of fine solid particles), a soft mist, an aerosol or a dry powder inhaler.

In one embodiment of the invention, a method of making the interferon lambda of the invention is provided.

The interferon lambda of the present invention may be prepared by a method comprising expressing a DNA sequence encoding the polypeptide in a host cell under conditions permitting expression of the interferon lambda peptide and then recovering the resulting peptide.

The medium used to culture the cells can be any conventional medium used to culture the host cells, such as minimal medium or complex medium containing suitable additives. Suitable media can be obtained commercially or prepared according to published procedures. The polypeptide produced by the host cell can then be recovered from the culture medium by conventional methods, for example, by precipitating the protein component of the supernatant or filtrate with a salt such as ammonium sulfate, and further purified by various chromatographic methods such as, for example, exchange chromatography, gel filtration chromatography, affinity chromatography, etc., depending on the kind of the desired peptide.

The coding DNA sequence described above may be inserted into any suitable vector. In general, the choice of vector will often depend on the host cell into which the vector is to be introduced, and thus, the vector may be an autonomously replicating vector, i.e., a vector which exists as an extrachromosomal entity, the replication of which is independent of chromosomal replication, e.g., a plasmid. Alternatively, the vector may be of a type which, when introduced into a host cell, is integrated into the host cell genome and replicated together with the chromosome(s) into which it has been integrated.

The vector is preferably an expression vector in which the DNA sequence encoding the peptide is operably linked to other segments of the DNA required for transcription, such as a promoter. Examples of promoters suitable for directing transcription of DNA encoding a peptide of the invention in a variety of host cells are well known in the art, see for example Sambrook, J, Fritsch, EF and maniotis, T, molecular cloning: a guide to the experimental work, Cold Spring Harbor Laboratory Press, New York, 1989.

The vector may also contain a selectable marker, e.g., a gene the gene product of which complements a defect in the host cell or which confers resistance to a drug, e.g., ampicillin, doxorubicin, tetracycline, chloramphenicol, neomycin, streptomycin, or methotrexate.

To introduce the expressed peptides of the invention into the secretory pathway of a host cell, a secretory signal sequence (also referred to as a leader sequence) may be provided in the recombinant vector. The secretory signal sequence is linked in the correct reading frame to the DNA sequence encoding the peptide. The secretion signal sequence is usually located 5' to the DNA sequence encoding the peptide. The secretory signal sequence may be one normally linked to the peptide, or may be derived from a gene encoding another secretory protein.

Methods for ligating the DNA sequence encoding the peptide of the present invention, the promoter and optionally the terminator and/or secretion signal peptide sequence, respectively, and inserting them into a suitable vector containing information necessary for replication are known to those skilled in the art.

The host cell into which the DNA sequence or recombinant vector is to be introduced may be any cell capable of producing the peptide of the invention, including bacterial, yeast, fungal and higher eukaryotic cells. Examples of suitable host cells well known and used by those skilled in the art include, but are not limited to: e.coli, Saccharomyces cerevisiae, or mammalian BHK or CHO cell lines.

For therapeutic use, the skilled artisan can readily determine the appropriate dosage, frequency of administration, and route of administration. Factors for making such determinations include, but are not limited to, the nature of the protein used, the condition to be treated, the potential patient compliance, the age, weight, sex, physical health, nutritional status, time of administration, metabolic rate, severity of the condition, and the judgment of the attending physician, e.g., a dosage of 1 μ g to 100mg, more preferably 1 μ g to 50mg, or 1 μ g to 10mg, or 10 μ g to 50mg, or 100 μ g to 50mg, or 1mg to 100mg, as will be appreciated by those skilled in the art, may be suitably administered depending on the actual condition.

In still another aspect, the invention also relates to a kit or kit comprising the pharmaceutical composition, formulation, medicament. In addition to the above-mentioned drugs or preparations, the kit or kit may further comprise other drugs, drug compounds or compositions which can be used in combination with the drug composition, preparation or medicament, for example, the other drugs, drug compounds or compositions may be selected from other coronavirus drugs, such as faviravir, chloroquine, oseltamivir, type I interferon, or other Chinese patent drugs.

The invention is further illustrated by the following examples, which, however, should not be construed as limiting the scope of protection of the present patent, the features disclosed in the foregoing description and in the following examples (individually and in any combination thereof), may be material for realising the invention in substantially different forms, in any combination thereof. In addition, the present invention incorporates publications which are intended to more clearly describe the invention, and which are incorporated herein by reference in their entirety as if reproduced in their entirety.

Drawings

FIG. 1 is a graph of the inhibition of new coronaviruses by interferon lambda 1.

FIG. 2 is a graph of the cytotoxicity of interferon lambda 1 against new coronaviruses.

FIG. 3 shows pathological changes in lesion of pulmonary bronchioles and pulmonary arterioles, wherein A is normal group; b, model group; c is DEX group; d, XW001-0.24 mg/kg; e, XW001-100 mg/kg.

FIG. 4 is pathological changes in alveolar injury, with normal groups; b, model group; c is DEX group; d, XW001-0.24 mg/kg; e, XW001-100 mg/kg.

FIG. 5 is a statistical chart of pathological injury scores of various parts of the mouse lung, which are sequentially from left to right, the total lung injury score, the total bronchiole and small pulmonary artery injury score and the total lung injury score.

The specific implementation mode is as follows:

example 1 was carried out: construction of interferon lambda expression engineering bacteria

A human interferon lambda 1/lambda 2/lambda 3/lambda 4 gene fragment (NM-172140.2/NM-172138.2/NM-172139.2) is obtained by a chemical synthesis mode, and is inserted into a prokaryotic expression plasmid pET-30a through Node I and Xho I sites and is sequenced for verification. The resulting expression plasmid was used for transformation assays.

BL21(DE3) competent cells (Invitrogen) were transformed with the plasmid containing the desired gene obtained above, and 50. mu.l of BL21 competent cells were thawed on an ice bath, the desired DNA was added thereto, shaken gently, and left in the ice bath for 30 minutes. Followed by heat shock in a water bath at 42 ℃ for 30 seconds, and then the centrifuge tube was quickly transferred to an ice bath for 2 minutes without shaking the centrifuge tube. The tubes were mixed with 500. mu.l of sterile LB medium (containing no antibiotics) and incubated at 37 ℃ and 180rpm for 1 hour to resuscitate the bacteria. Mu.l of the transformed competent cells were pipetted onto a plate of LB agar medium containing kanamycin resistance and the cells were spread out evenly. The plate was placed at 37 ℃ until the liquid was absorbed, inverted and incubated overnight at 37 ℃. The following day, single colonies in the transformation plate were picked using an inoculating loop and inoculated in 15ml of sterile LB medium (containing antibiotics) and cultured overnight at 30 ℃.

Example 2 was carried out: expression and purification of interferon lambda

To 50ml of LB medium was added 50. mu.l of a bacterial solution (a bacterial solution expressing interferon. lambda.) and 50. mu.l of kanamycin, and after mixing, the mixture was placed in a 30 ℃ constant temperature shaker and inoculated overnight. 10ml of overnight inoculated broth was added to 1000ml of LB medium, together with 1000. mu.l of kanamycin. Shaking, culturing in a shaker at 37 deg.C at 200rpm until the OD600 of the bacterial liquid is 0.4-0.6, adding 0.5mM IPTG for induction, culturing for 4 hr, and collecting thallus. The expressed IL29 mutant accounts for about 30-50% of the total protein of the thallus and exists mainly in the form of inclusion body.

The fermented cells were washed 3 times with TE (10mmol/L Tris-HCl,1mmol/L EDTA, pH 6.5) solution (m: V ═ 1:10), then homogenized at 60MPa and disrupted, and the disrupted ratio was examined under a microscope. When the thallus breakage rate is about 95% (about 2-3 times) centrifuging at 8000rpm for 15min, and collecting broken thallus precipitate. The disrupted cell pellet was placed in a beaker, and an inclusion body washing solution (10mM Tris-HCl +1mM EDTA + 0.5% Triton-X100, pH 6.5, m: V: 1:10) was added thereto, and the mixture was stirred in a magnetic stirrer for 30min and washed 3 to 5 times. (see figure 1 for inclusion body SDS-PAGE) inclusion bodies were lysed with inclusion body lysates (7M guanidine hydrochloride +50mM Tris-HCl +10mM DTT, pH 6.5, M: V ═ 1:10), stirred at room temperature overnight. The cleaved protein was slowly added to a renaturation solution (100mM Tris-HCl, 0.5M Arginine, 0.5% PEG3350(M: V), 2mM GSH:0.5mM GSSG, pH 8.5) to a final protein concentration of 0.2mg/ml, and stirred at room temperature overnight.

The renaturation solution is centrifuged for 5min at 8000rpm, and the supernatant is collected. Using an ultrafiltration membrane pack having a pore size of 10kDa, the ultrafiltration membrane was equilibrated with 20mM phosphate buffer solution pH7.0, and then 1L of the supernatant was concentrated 10-fold. The concentrate was diluted with 5-fold volume of water for injection, and 600ml of a sample solution was finally collected. Loading the concentrated solution to a 20mM phosphate buffer solution, washing with pH7.0, loading, eluting with 50mmol/LTris-HCl, pH8.5,0.15mol/LNaCl, and collecting the peak components;

sepharose FF packed column, 20mmol/L phosphate, pH7.4,0.05mol/L NaCl equilibration, load until the detector baseline is stable. Washing with 20mmol/LPB, pH7.4 and 0.2mol/L NaCl solution, and collecting the eluted peak components.

And G25Media is filled into a column, a 20mM disodium hydrogen phosphate buffer solution is balanced and loaded, elution peak components are collected, and the purity of the finally obtained interferon lambda is more than 95% after the purification process.

Example 3 of implementation: novel coronavirus in vitro drug effect

Cell: VeroE6 cells, preserved by the institute for pathogenic bacteria, institute for medical laboratory, Chinese academy of medical sciences.

Virus: 2019-nCoV with the titer of 10⁵TCID₅₀Perml, preserved at-80 deg.C by the institute of laboratory animal medicine, institute of Chinese academy of medicine and sciences. Using a Virus titre of 100TCID₅₀。

Sterile 96-well culture plate, 200. mu.l of 5X 10 concentration per well⁴cell/ml Vero E6 cells, 5% CO at 37 ℃₂Culturing for 24 hours;

(2) diluting the tested medicine into 2 concentrations, each concentration is 5 multiple wells, and 100 mu l of the diluted medicine is added into each well for 24 hours;

(3) add 100. mu.l of 100TCID per well₅₀A virus;

(4) simultaneously setting a cell control, a blank control (solvent control) and a virus control (negative control);

(5) cells at 37 ℃ and 5% CO₂Incubating in an incubator for 4-5 days;

(6) cytopathic effects (CPE) were observed under light microscopy, with complete lesions recorded as "++++", 75% lesions recorded as "++++", 50% lesions recorded as "+++", 25% lesions recorded as "+", and no lesions recorded as "-".

IFN lambda 1 anti-novel coronavirus 2019-nCoV effect

And (4) conclusion: under the experimental conditions IFN lambda 1/lambda 2/lambda 3/lambda 4 at a concentration of 10ng/ml (0.5nmol/L) protected 100% cells from infection by the new coronavirus, showing very good therapeutic potential.

Example 4 of implementation: novel coronavirus in vitro drug effect

And (3) experimental operation:

cell: VeroE6 cells, interferon lambda 1/lambda 2/lambda 3.

(1) Sterile 96-well culture plate, 200. mu.l of 5X 10 concentration per well⁴cell/ml Vero E6 cells, 5% CO at 37 ℃₂Culturing for 24 hours;

(2) diluting the tested medicine into 9 concentrations, each concentration is 2 multiple wells, and 100 mu l of the diluted medicine is added into each well for 24 hours;

(3) add 100. mu.l of 100TCID per well₅₀A virus;

(5) cells at 37 ℃ and 5% CO₂Incubating in an incubator for 4-5 days; optical systemObserving cytopathic effect (CPE) under microscope, culturing until the pathological change of virus control group reaches 4+, and calculating half effective concentration EC by Reed-Muenc method₅₀。

The EC50 of interferon lambda 1 against the novel coronavirus was 2.087ng/ml as seen in antiviral experiments, while cytotoxicity experiments showed IC₅₀Greater than 200. mu.g/ml. SI is greater than 100,000. There is a very large security window.

Example 5

According to the results of the related studies, patients with new coronavirus infection are responsible for respiratory distress syndrome (ARDS) by the cytokine storm caused after the viral infection, and thus the occurrence of these complications can be controlled during the treatment.

Experimental animals: male C57 mice, 18-22g, 30, Shanghai Ling Biotech Co., Ltd

Establishing a model:

after the animal is anesthetized, the skin is cut open to expose the trachea; LPS solution (50 ul/mouse, 0.3mg/kg) was slowly injected through trachea to replicate the acute lung injury model; animal body weight and health were monitored for a total of 24 hours from the start of modeling to the end of the experiment.

Experiment design:

the results are shown in FIGS. 3 to 5

The results show that: research in an LPS-induced mouse acute lung injury model shows that in histological evaluation, compared with a model group, an interferon administration group has significant curative effects of relieving lung injury and inflammatory reaction, and compared with the model group, the difference of a treatment group has significance; in the aspects of reducing the total pulmonary alveolar injury score and the total lung injury score, the interferon lambda 1 atomizing administration mode is obviously superior to the interferon alpha 2b group, and the interferon lambda 1 is proved to better control the cytokine storm caused in the treatment process and relieve the respiratory distress syndrome (ARDS) caused by the cytokine storm in the process of treating patients infected by new coronavirus with interferon.

Sequence listing

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<120> use of interferon lambda in the treatment of infections with novel coronaviruses (2019-nCoV)

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Claims

1. Use of interferon lambda for the preparation of a medicament for the prevention and/or treatment of a novel coronavirus infection.

2. Use of a pharmaceutical composition for the manufacture of a medicament for the prevention and/or treatment of a novel coronavirus infection, wherein the composition comprises interferon lambda, and a pharmaceutically acceptable adjuvant or excipient.

3. The use of claim 1 or 2, wherein the interferon λ is interferon λ 1, interferon λ 2, interferon λ 3 or interferon λ 4.

4. The use of claim 1 or 2, wherein the interferon λ reduces the level of virus in a patient infected with a new coronavirus.

5. The use of claim 1 or 2, wherein the interferon λ reduces damage to airway epithelial cells caused by neutrophils.

6. The use according to claim 1 or 2 wherein the interferon λ alleviates respiratory distress syndrome caused by a new coronavirus infection.

7. An article of manufacture comprising a container having the pharmaceutical composition of claim 2 therein and a package insert, wherein the package insert carries instructions for use of the pharmaceutical composition.

8. The article of manufacture of claim 7, further comprising a container containing an additional pharmaceutical agent.

9. The article of manufacture of claim 8, wherein the additional agent is an additional agent for the treatment of a coronavirus.