WO2022014813A1

WO2022014813A1 - Pharmaceutical composition for preventing or treating sars-cov-2 infection

Info

Publication number: WO2022014813A1
Application number: PCT/KR2021/002355
Authority: WO
Inventors: Min-Hyo Ki; Moon-Jung BACK; Jang-Hyun Lee; Jong-Hwan Lee; Hyeong-Woo MOON
Original assignee: Samjin Pharmaceutical Co Ltd
Current assignee: Samjin Pharmaceutical Co Ltd
Priority date: 2020-07-15
Filing date: 2021-02-25
Publication date: 2022-01-20
Anticipated expiration: 2023-01-15

Abstract

The present disclosure relates to a pharmaceutical composition for preventing or treating SARS-CoV-2 infection, and more particularly to, a pharmaceutical composition for preventing or treating SARS-CoV-2 infection, containing nelfinavir expressed by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human; a use of nelfinavir, expressed by the following chemical formula 1, or a pharmaceutically acceptable salt thereof for preventing or treating and for preparing a medicament for preventing or treating SARS-CoV-2 infection using the nelfinavir or the pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human; and a method for preventing or treating SARS-CoV-2 infection, including administering nelfinavir, expressed by the following chemical formula 1, or a pharmaceutically acceptable salt thereof to a subject, requiring the nelfinavir or the pharmaceutically acceptable salt thereof, as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

Description

PHARMACEUTICAL COMPOSITION FOR PREVENTING OR TREATING SARS-COV-2 INFECTION

The present disclosure relates to a pharmaceutical composition for preventing SARS-CoV-2 infectious diseases or treating diseases caused by SARS-CoV-2 infection.

Coronavirus is an RNA virus belonging to the subfamily Coronavirinae in family Coronaviridae, which is enveloped virus with a positive ssRNA and a nucleocapsid of helical symmetry. Coronaviruses are named after the Latin word corona, referring to crown, and have crown-like spike proteins, club-shaped projections, protruding from the surfaces thereof, resembling the sun's corona.

In 1937, the first coronavirus was discovered in chickens. Coronaviruses, found in various animals such as bats, birds, cats, dogs, cows, pigs, mice, and the like, are classified into four groups (alpha-, beta-, gamma-, and delta coronaviruses). The alpha-coronaviruses and beta-coronaviruses mainly infect mammals, and the gamma-coronaviruses and delta-coronaviruses can be identified in birds. Coronaviruses are reported to cause various diseases such as stomach disorder and respiratory diseases in animals.

Human coronaviruses infecting humans, reported to Centers for Disease Control and Prevention (CDC), include HCoV-229E and HCoV-OC43 discovered in the 1960s, HCoV-NL63 (2004) and HCoV-HKU1 (2005) discovered after the severe acute respiratory syndrome (SARS, discovered in 2003) coronavirus (SARS-CoV) pandemic, and middle east respiratory syndrome (MERS) coronavirus (MERS-CoV) having severe respiratory symptoms such as high fever, cough, and respiratory distress, similar to those of SARS, identified first in Saudi Arabia in September 2012. Recently, the SARS-coronavirus (SARS-CoV-2), which originated in Wuhan in December 2019 and has spread worldwide, was reported to be the seventh strain of human infectious coronavirus, different from the existing six strains of coronavirus.

Coronavirus disease 2019 (hereinafter, interchangeable referred to as "COVID-19") is a respiratory infection disease caused by SARS-CoV-2, the new type of coronavirus (hereinafter, interchangeably referred to as "the novel coronavirus"). COVID-19 may be spread through droplets from coughing or sneezing of an infected person which permeate into mucous members of eyes, nose, mouth, and the like, or may be spread by touching eyes, nose, and mouth after contact with a contaminated object or hand. In addition, the possibility of airborne transmission of COVID-19 is under discussion, and infectibility of COVID-19 is significantly high. When a person is infected with the novel coronavirus, the person shows various degrees of symptoms from mild symptoms to severe symptoms and shows main symptoms such as fever, cough, respiratory distress, pneumonia, diarrhea, abdominal pain, abdominal discomfort, or the like, after an incubation period estimated to be two to fourteen days. A symptom such as a sore throat, blood stasis, or nausea is often exhibited. Furthermore, cases of asymptomatic infection have been reported. COVID-19 has a significantly high fatality rate (more than 340,000 people were killed around the world as of May 2020), but development of effective preventive or therapeutic medicines remains in progress. Accordingly, there is an urgent need for the development of drugs to prevent or treat COVID-19.

Hereinafter, drug properties of nelfinavir to be used and background arts, in which access possibility of nelfinavir for COVID-19 is mentioned, will be described below.

Nelfinavir is a drug developed as an anti-AIDS drug, and is developed to be used to treat AIDS infections by being taken in a dose of 2,500 mg or 2,250 mg per day for an adult human. The document regarding side effects of nelfinavir (Pharmacotherapy, 2004, 24(6), 727-735) states that when nelfinavir is taken at a dose of 2,500 mg or 2,250 mg per day for an adult human as a treatment of AIDS, about 49% of people showed a symptom of diarrhea as a side effect of nelfinavir and about 33% of people should be medically treated for a symptom of diarrhea.

The document regarding cytotoxicity of nelfinavir (Autophagy, 2008, 4(1), 107-109) states that nelfinavir amplifies endoplasmic reticulum stress and increases autophagy to increase cellular death caused by apoptosis and non-apoptosis. Nelfinavir has been developed as an anticancer drug based on a multifactorial cytotoxicity effect using such autophagy or the like, and clinical trials for anticancer drugs have been made within the range of a dose of anti-AIDS drug. This means that when nelfinavir is taken within the range of a dose of an anti-AIDS drug, blood concentration may already reach a value at which cytotoxicity required for anticancer treatment is induced. In addition, the document regarding a clinical trial for an anticancer drug of nelfinavir (Head Neck. 2015, 37(5), 722-726) states that among fifteen adenoid cystic carcinoma patients who administered nelfinavir at a dose for anti-AIDS, five patients stopped a clinical trial due to side effects of the drug. Therefore, it can be seen that there are many cases of side effects, beyond the anticancer effect, even in anticancer clinical trials.

In US Patent Registration No. 7,495,011, it is set forth that nelfinavir is used for SARS-Coronavirus (SARS-CoV). However, SARS-CoV and SARS-CoV-2 are genetically different from each other and, even in an asymptomatic state, SARS-CoV-2 has a higher viral load than SARS-CoV to show infectivity, and thus, has a significantly high transmission rate, which is different in epidemiological aspect. In addition, considering that many substances and drugs including chloroquine, and the like, reported to be effective on SARS-CoV, are not effective on SARS-CoV-2, it is significantly unpredictable as to whether nelfinavir have therapeutic efficacy against SARS-CoV-2, genetically different from SARS-CoV, and as to a dosage for obtaining the treatment effect against COVID-19, through US Patent Registration No. 7,495,011.

Some documents (J Med Virol., 2020, 1-9; DOI: 10.26434/chemrxiv.12039888.v1; DOI: 10.1101/2020.04.06.026476; DOI: 10.1101/2020.04.14.039925; DOI: 10.1101/2020.06.22.165712) disclose IC ₅₀ in the range of 0.77 to 10 μM, in relation to an SARS-CoV-2 inhibition effect of nelfinavir using Vero cell line, a monkey kidney cell line, or A549 cell line modified from human lung cells. However, the above documents confirm only a little possibility because the effect on SARS-CoV-2 was evaluated by expressing artificial proteins in cells which did not express or hardly expressed proteins such as ACE2 and TMPRSS2 known to be the major factors of human infection routes of SARS-CoV-2. Therefore, the above documents fail to determine whether there is an actual treatment effect for SARS-CoV-2 and to find an effective dose.

In addition, some literature (DOI: 10.21203/rs.3.rs-20948/v1; J. Phys. Chem. Lett., 2020, 11, 4413-4420) disclose possibility of SARS-CoV-2 inhibition by nelfinavir through computer simulations. However, since the simulations were not actual tests, the above documents fail to precisely predict whether there is an actual treatment effect for SARS-CoV-2 and fail to precisely predict an effective dose.

One clinic example report (DOI: 10.21203/rs.3.rs-27346/v1) discloses a case report in which nelfinavir was applied to six COVID-19 patients in the situation in which many drugs have been tested in humans due to the COVID-19 pandemic. According to the case report, two patients with predominant gastrointestinal symptoms such as diarrhea stopped taking the drug due to dehydration or showed only little improvement in the symptoms. Therefore, there was an obvious limitation in applying the same dose as a dose of nelfinavir used as an anti-AIDS drug for treatment of COVID-19 because of the concern about side effects of nelfinavir such as diarrhea. The case report discloses that among four patients who administered a dose of nelfinavir used as an anti-AIDS drug, three patients responded to treatment, but discloses only results obtained when the three patients took a combination of nelfinavir and baloxavir, or had already been taking hydroxychloroquine and baloxavir. Therefore, the case report is inappropriate as base data of a clinical treatment effect of nelfinavir alone, and it is unpredictable as to whether nelfinavir has a treatment effect for SARS-CoV-2, or the like, without occurrence of side effects and as to a dosage of nelfinavir for obtaining effective treatment effect.

Accordingly, the present disclosure provides a low dose of a nelfinavir composition, capable of obtaining a treatment effect for COVID-19 while reducing side effects, and uses thereof.

An aspect of the present disclosure is to provide a pharmaceutical composition for preventing or treating SARS-CoV-2 infection using nelfinavir or a pharmaceutically acceptable salt thereof at a low dose.

Another aspect of the present disclosure is to provide a novel use of a low dose of nelfinavir or a pharmaceutically acceptable salt thereof.

Another aspect of the present disclosure is to provide a method for preventing or treating SARS-CoV-2 infection using a low dose of nelfinavir or a pharmaceutically acceptable salt thereof.

Example embodiments of the present disclosure provide a pharmaceutical composition for preventing or treating SARS-CoV-2 infection, containing nelfinavir expressed by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

[Chemical Formula 1]

Example embodiments of the present disclosure provide a use of nelfinavir or a pharmaceutically acceptable salt thereof for preventing or treating SARS-CoV-2 infection using the nelfinavir or the pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

Example embodiments of the present disclosure provide a use of nelfinavir or a pharmaceutically acceptable salt thereof for preparing a medicament for preventing or treating SARS-CoV-2 infection using the nelfinavir or the pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

Example embodiments of the present disclosure provide a method for preventing or treating SARS-CoV-2 infection, including administering nelfinavir or a pharmaceutically acceptable salt thereof to a subject in need of treatment, as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

According to the present disclosure, a composition, containing nelfinavir for preventing or treating SARS-CoV-2 or a pharmaceutically acceptable salt thereof at a low dose, and uses of the composition are provided. In particular, a low dose of nelfinavir or a pharmaceutically acceptable salt thereof may significantly reduce side effects while being administered at a lower dose than a common dose of an existing anti-AIDS drug or anticancer drug and, and thus, are expected to be useful in treating SARS-CoV-2 infection, in detail, COVID-19 because side effects unfavorable to treatment, such as gastrointestinal side effects, are reduced or are not accompanied and even gastrointestinal symptoms of COVID-19 are not exacerbated.

FIG. 1 illustrates a dose-response curve of a compound against the expression level of SARS-CoV-2.

Hereinafter, example embodiments of the present disclosure will be described. However, the embodiments of the present disclosure may be modified to have various other forms, and the scope of the present disclosure is not limited to the embodiments described below.

The present disclosure is based on expression of a significant treatment effect and discovery the possibility of reducing side effects when nelfinavir, known as a conventional anti-AIDS drug, or a pharmaceutically acceptable salt thereof are used. Accordingly, the present disclosure provides a pharmaceutical composition, containing nelfinavir for preventing or treating SARS-CoV-2 infection accompanying COVID-19 at a low dose or a pharmaceutically acceptable salt thereof, novel uses of the nelfinavir and the pharmaceutically acceptable salt thereof, and a method of preventing or treating SARS-CoV-2 infection using the nelfinavir and the pharmaceutically acceptable salt thereof.

Specifically, the present disclosure provides a pharmaceutical composition for preventing or treating SARS-CoV-2 infection, containing and using nelfinavir expressed by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg to 1,500 mg per day for an adult human.

[Chemical Formula 1]

Since the pharmaceutical composition of the present disclosure contains the nelfinavir or a pharmaceutically acceptable salt thereof with low dose, gastrointestinal side effects may be reduced or may not be accompanied, and gastrointestinal symptoms may not be exacerbated. As described above, the pharmaceutical composition of the present disclosure may use low dose of nelfinavir to reduce or prevent side effects in patients who have no gastrointestinal symptoms; and/or to prevent exacerbation of gastrointestinal symptoms and to express even effects of treating and relieving gastrointestinal symptoms of patients who have the gastrointestinal symptoms.

The present disclosure provides uses of nelfinavir or a pharmaceutically acceptable salt thereof for preventing or treating SARS-CoV-2 infection using nelfinavir expressed by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg to 1,500 mg per day for an adult human.

In addition, the present disclosure nelfinavir or a pharmaceutically acceptable salt thereof for preparing a medicine for preventing or treating SARS-CoV-2 infection using nelfinavir expressed by the above chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg to 1,500 mg per day for an adult human.

[Chemical Formula 1]

With the low dose of nelfinavir or the pharmaceutically acceptable salt thereof, gastrointestinal side effects may be reduced or may not be accompanied, and gastrointestinal symptoms may not be exacerbated.

Hereinafter, components and detailed contents of the present disclosure will be described.

The name of a compound in the chemical formula 1 is ([3S-[2(2S*, 3S*), 3α,4aβ,8aβ]]-N-(1,1-dimethylethyl)decahydro-2-[2-hydroxy-3-[(3-hydroxy-2-methylbenzoyl)amino]-4-(phenylthio)butyl]-3- isoquinoline carboxamide), and will be referred to interchangeably with nelfinavir.

In the present specification, the term "active ingredient" refers to target activation alone, or an ingredient effective in recovering a disease defined as "SARS-CoV-2 infection" (relieving a symptom of the disease), treating the disease, and preventing the disease (inhibiting or delaying occurrence of the disease) and may be chemically, in detail, a free base not containing salt or hydrate.

In the present specification, the term "prevention" refers to all actions for inhibiting infection by the novel coronavirus or delaying occurrence thereof, may include reduction in a hazard ratio of infection.

In the present specification, the term "treatment" refers to all actions for recovering, fully covering, or beneficially changing at least a portion or entire symptom of COVID-19, caused by novel coronavirus infection, by the present disclosure.

In the present disclosure, the term "coronavirus" refers to single-stranded RNA coronavirus belonging to Coronaviridae family and Coronavirinae subfamily, and may be alpha coronavirus or beta coronavirus, in detail, beta coronavirus. For example, the term "coronavirus" may include human coronavirus, infecting human, and variants thereof. The coronavirus, to which the present disclosure may be applied, may be, in detail, SARS-CoV-2. For example, a pharmaceutical composition of the present disclosure may be used to prevent or treat COVID-19 caused by SARS-CoV-2.

COVID-19 may accompany symptoms which may be accompanied by malaise and/or fever (37.5 degrees) and/or redness after an incubation period of 1 to 14 days. The symptoms may include at least one selected from the group consisting of loss of at least one of taste and smell, respiratory symptom such as phlegm, nasal stuffiness, coughing, dyspnea, sore throat, headache, gastrointestinal symptoms including diarrhea and/or abdominal pain and abdominal discomfort, hemoptysis, nausea, body aches, muscular pain, joint pain, chest pain, chest tightness, chill, vertigo, lymphopenia, hemocoagulation, hematologic symptoms such as an increase in inflammatory index, dysphrasia, dyscinesia, mental aberration, anxiety, depression, somnipathy, stroke, delirium, brain injury, nerve injury, liver injury, kidney injury, skin rash, skin discoloration, acute myocardial infarction, myocarditis, cardiac insufficiency, conjunctivitis, upper respiratory tract infection, lower respiratory tract infection, respiratory infection, pneumonia, eye infection, kidney infection, liver infection, skin infection, nervous system infection, cardiovascular infection, and the like, but is not limitedn discoloration, acute myocardial infarction, myocarditis, cardiac insufficiency, and red eye, which are caused by SARS-CoV-2. In addition, the corona virus disease-19 may be at least one selected from the group consisting of upper respiratory tract infection of SARS-CoV-2, lower respiratory tract infection of SARS-CoV-2, respiratory organ infection of SARS-CoV-2, gastrointestinal tract infection of SARS-CoV-2, pneumonia caused by SARS-CoV-2, ophthalmic infection of SARS-CoV-2, skin infection of SARS-CoV-2, nervous system infection of SARS-CoV-2, cardiovascular system infection of SARS-CoV-2, kidney infection of SARS-CoV-2, liver infection of SARS-CoV-2, and a blood corpuscle infection of SARS-CoV-2.

The term "respiratory organ" refers to an organ involved in inh thereto. In addition, it will be understood that the above-described symptoms includes all symptoms caused by SARS-CoV-2 infection. Furthermore, there is an asymptomatic case. Therefore, COVID-19 caused by SARS-CoV-2, to which the present disclosure is applied, is intended to include not only a case in which at least one of the symptoms is expressed but also an asymptomatic case in which no symptom is expressed.

The "subject" or "patient", to which a pharmaceutical composition for preventing or treating SARS-CoV-22 infection of the present disclosure is applied, may be a subject who exhibits the symptom, not limited to the above-mentioned symptoms, caused by SARS-CoV-2 infection and/or at least one of the other symptoms, a subject who has an asymptomatic virus, or a subject which is concerned about coronavirus infection. For example, "subject" may include any subject which participated in a clinical study trial showing no clinical finding for any disease, any subject which participated in an epidemiological study, or any subject used as a control group.

Specifically, the "subject" or "patent" may be, for example, mammals and may be, in detail, any subject selected from the group consisting of humans, cow, dog, cat, pig, goat, guinea pig, rabbit, chicken, and the like and required to be treated.

More specifically, COVID-19 may accompany at least one of the symptoms such as fever, redness, loss of at least one of taste and smell, phlegm, coughing, sore throat, headache, diarrhea, abdominal pain, abdominal discomfort, hemoptysis, nausea, dyspnea, nasal stuffiness, body aches, muscular pain, joint pain, chest pain, chest tightness, fatigue, chill, vertigo, lymphopenia, hemocoagulation, dysphrasia, dyscinesia, mental aberration, anxiety, depression, somnipathy, stroke, delirium, brain injury, nerve injury, liver injury, kidney injury, skin rash, skialing and exhaling, and may include an upper respiratory tract and a lower respiratory tract. The "upper respiratory tract" includes mouth, nose, sinuses, middle ear, throat, larynx, and trachea, and the "lower respiratory tract" includes bronchial tube and lungs (bronchi, bronchioles, and alveoli) as well as interstitial tissue of the lungs.

The term "gastrointestinal tract" refers to a canal from the mouth to the anus, including mouth, esophagus, stomach and intestines.

The term "nervous system" refers to a system responding to, transmitting, integrating, and determining stimuli and transmitting commands. The "nervous system" includes brains including cerebrum, cerebellum, diencephalon, mesencephalon, and medulla oblongata, a central nervous system including spinal cord, a peripheral nervous system, and an autonomic nervous system.

The term "cardiovascular system" may be used interchangeably with the term "circulatory system", and may include heart and blood vessels.

The term "pharmaceutically acceptable salt" refers to a formulation of a compound which does not cause serious stimulus in an organism, to which the compound is administered, and does not damage biological activity and physical properties of the compound. The pharmaceutical acceptable salt includes acid addition salt prepared by acid preparing non-toxic acid addition salt containing pharmaceutically acceptable anions, for example, inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrobromic acid (bromic acid), hydroiodic acid (iodic acid), perchloric acid, or tartaric acid, organic carboxylic acid such as succinic acid, oxalic acid, tartaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid, tartaric acid, formic acid, citric acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid, benzoic acid, lactic acid, fumaric acid, maleic acid, salicylic acid, or sulfonic acid such as methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, or the like. In the present disclosure, the pharmaceutically acceptable salt thereof may be selected from, in detail, mesilate salt, methanesulfonic acid, and benzenesulfonic acid. The compound according to the present disclosure may be converted into salt thereof using a conventional method.

In conventional research into a dose of nelfinavir, the dose has been estimated by expressing artificial protein in cells which do not express proteins such as ACE2 and TMPRSS2 known as major factors in human infection routes of SARS-CoV-2 and disclosed only IC ₅₀ in the range of 0.77 μM to 10 μM. Therefore, the conventional research fails to derive an effective low dose, a target of the present disclosure. Meanwhile, in the present disclosure, IC ₅₀ of nelfinavir was confirmed to be 0.01 μM using Calu-3 cell line, a human lung cell line in which proteins such as ACE2 and TMPRSS2 important for human cell infection of SARS-CoV-2 were expressed well, and a low effective dose of the present disclosure was derived based on the confirmation.

The term "low dose" refers to a dose below a usual dose (a common dose), for example, a dose lower than or equal to a usual dose when nelfinavir is used as an anti-AIDS medicine which may be calculated as a dose per day (a daily dose). For example, even when divided into two or three parts to be taken twice or three times a day, a daily total dose of nelfinavir is calculated to be lower than a daily dose of nelfinavir used as an anti-AIDS medicine. In this case, the dose is referred to as "low dose."

According to a treatment when nelfinavir is used as an anti-AIDS medicine, it is specified that 1,250 mg of nelfinavir is taken twice a day for an adult human or 750 mg of nelfinavir is taken three times a day for an adult human. That is, a total daily dose of nelfinavir is 2,500 mg or 2,250 mg. Unlike the known total daily dose of nelfinavir, in the present disclosure, a dose of nelfinavir or a pharmaceutically acceptable salt thereof, based on a usual dose for an adult human, is 100 mg/day or more to 1,500 mg/day or less, for example, 100 mg/day or more to less than 1,500 mg/day, in detail, for example, 100 mg/day, or 250 mg/day, 300 mg/day, 500 mg/day, 600 mg/day, 750 mg/day, 900 mg/day, 1,000 mg/day, 1,200 mg/day, 1,250 mg/day, 1,400 mg/day, and 1,500 mg/day, and is intended to include numerical ranges of all combinations which may be derived within the above range.

The present disclosure is aimed at administering nelfinavir or a pharmaceutically acceptable salt thereof at a dose of 100 mg/day to 1,500 mg/day, and the nelfinavir, or the pharmaceutically acceptable salt thereof, or a pharmaceutical composition including the same can be administered once a day or can be divided into two or four parts to be administered twice or four times a day. In the present specification, each numerical range is intended to include the numerical range of all combinations which may be derived within the range, and is not limited as examples of the above-mentioned numerical range.

For example, in the present disclosure, when nelfinavir is divided into two parts to be administered twice a day, 50 mg to 750 mg of the nelfinavir may be administered at a single dose. When nelfinavir is divided into three parts to be administered three times a day, 33.3 mg to 500 mg of the nelfinavir may be administered at a single dose. In the present disclosure, such a low dose may be effectively used to relieve COVID-19 symptoms because nelfinavir may exhibit efficacy for preventing or treating SARS-CoV-2 infection at a low dose while reducing cytotoxicity or side effects which may occur in a usual dose as an existing anti-AIDS medicine.

The term "effective dose" may be used interchangeably with the "therapeutic dose", and is 100 mg/day or more to 1,500 mg/day or less for an adult human. The term "effective dose" refers to a dose of an active ingredient which may exhibit intended medical and pharmacological effects, such as an effect of relieving SARS-CoV-2 infection during an administration period based on suggestions of medical experts, for mammals, in detail, people, an application target of the present disclosure. Such an effective dose is based on an adult having a weight of about 60 kg.

In the case of COVID-19, since primary infection occurs from a wide part of mucous membrane, various inflammatory responses to bronchial tube, lung, gastrointestinal tract, and the like, mucosal tissues exposed outwardly, are intensified. Accordingly, gastrointestinal symptoms such as abdominal pain, abdominal discomfort, and diarrhea as well as respiratory symptoms such as cough, dyspnea, and pneumonia are accompanied, so that side effects of nelfinavir may be a serious issue in COVID-19.

Up to the present, although nelfinavir has various side effects, 1,250 mg of nelfinavir has been inevitably administered twice a day, or 750 mg of nelfinavir has been inevitably administered three times a day (i.e., a daily dose of 2,500 mg or 2,250 mg has been administered) as an anti-AIDS medicine or an anticancer medicine to express efficacy of nelfinavir. When the conventional dose of nelfinavir is administered, nelfinavir is known to cause typical side effects such as diarrhea, fatigue, headache, and the like. In particular, it was reported that diarrhea symptoms were shown in about 49.4% of total patients, and 33% of total patients needed treatment for diarrhea symptoms (Pharmacotherapy 24(6), 727-35, 2004).

On the other hand, it was reported that gastrointestinal symptoms such as abdominal pain, abdominal discomfort, and diarrhea caused by COVID-19 were shown in about 61% of total COVID-19 patients (Gut 69(6), 997-1001, 2020). In addition, it was reported that among COVID-19 patients, patients having gastrointestinal symptoms had 50% higher probability of being severely ill than patients having no gastrointestinal symptoms (Gut gutjnl-2020-321751, 2020). Although an existing dose of nelfinavir has antiviral effects on SARS-CoV-2, side effects of nelfinavir itself may cause symptoms of COVID-19 patents, in particular, gastrointestinal symptoms, to be exacerbated or become more severe.

However, the present inventors unexpectedly found that nelfinavir is able to achieve sufficient medicinal effects at a low dose. Therefore, limiting the dose of nelfinavir to a maximum of 1,500 mg or less a day could be applied to the treatment of COVID-19 so that the side effects of nelfinavir, known to comprise the above-described gastrointestinal side effects, are not accompanied or are significantly reduced and gastrointestinal symptoms of COVID-19 are not exacerbated.

In particular, in embodiments of the present disclosure, a significantly excellent SARS-CoV-2 infection inhibition effect of nelfinavir having IC ₅₀ with a level of 0.01 μM in Calu-3 cells, the human lung cells, was confirmed. Accordingly, in order to exhibit treatment effectiveness most appropriate to COVID-19 patients who show various symptoms including gastrointestinal inflammation which occur simultaneously, a daily dose of nelfinavir should be a low dose of 1,500 mg or less to significantly reduce side effects of a drug and to optimize an effect of the drug.

In addition, according to embodiments of the present disclosure, it was confirmed that nelfinavir had excellent IC ₅₀ of 0.01 μM (5.7 ng/mL) in Calu-3 cells, the human lung cells, to exhibit a significantly excellent inhibition effect against SARS-CoV-2. Based on the data and the Nelfinavir PK Document (Ther Drug Monit, Vol. 23, No. 4, 2001), a dose of nelfinavir to be maintained at 0.1 μM (57 ng/mL), 10 times the IC ₅₀ of the nelfinavir, or more during administration of the nelfinavir may be calculated. The calculated dose of nelfinavir corresponds to a dose when about 40 mg is taken twice a day and a dose when about 20 mg is taken three times a day.

For example, it means that when 40 mg of nelfinavir is administered twice a day or 20 mg of nelfinavir is administered three times a day, even lowest blood concentration immediately before next administration of nelfinavir after administration of nelfinavir may be maintained at blood concentration corresponding to 10 times or higher than the IC ₅₀ of nelfinavir against SARS-CoV-2. Since the above doses are respectively converted into daily doses of 80 mg and 60 mg, about 100 mg or more should be administered at a daily dose to secure a minimum effect of nelfinavir against SARS-CoV-2. For the reference, the calculation of 10 times the IC ₅₀ takes a margin of safety into account, considering a clinical individual difference.

Conventionally, nelfinavir has been concomitantly administered with other antiviral agents for the purpose of AIDS treatment or anti-cancer drug. However, it is not preferable that nelfinavir is concomitantly administered with other antiviral agents, other than drugs proved to have superior antiviral effects for SARS-CoV-2, for the purpose of COVID-19 treatment because side effects caused by nelfinavir and/or other antiviral agents may occur or may be exacerbated. Other antiviral agents, which is not preferable to be concomitantly administered with nelfinavir, may be saquinavir, lopinavir, ritonavir, amprenavir, indinavir, atazanavir, fosamprenavir, darunavir, and the like, as HIV protease inhibitors, and tenofovir, lamivudine, zidovudine), dianosine, emtricitabine, zalcitabine, stavudine, abacavir, and the like, as reverse transcriptase inhibitors. When such antiviral agents are concomitantly administered, various side effects such as diarrhea, fatigue, and headache may be accompanied. In particular, HIV protease inhibitors may cause gastrointestinal side effects such as severe diarrhea, and the like.

Accordingly, since concomitant administration of nelfinavir with HIV protease inhibitors and/or reverse transcriptase inhibitors may cause or exacerbate side effects such as diarrhea, nelfinavir should not be concomitantly administered with these medicaments in the case of treatment of COVID-19 patients.

However, the present disclosure is not limited thereto, and antiviral drugs having superior inhibitory effects on SARS-CoV-2 may be concomitantly administered as long as they do not cause side effects.

Based on the excellent effectiveness of nelfinavir against COVID-19 and PK characteristics of a significantly high C _trough, nelfinavir needs to be administered at a daily dose from 100 mg or more to 1,500 mg or less for an adult human to clinically apply the nelfinavir to treatment of COVID-19, in overall consideration of typical side effects and high cytotoxicity of nelfinavir and a wide range of mucosal side inflammatory symptoms such as gastrointestinal tract of COVID-19. Accordingly, the present disclosure is aimed at administering nelfinavir at a low dose of 100 mg/day to 1,500 mg/day for an adult human, rather than an existing high dose of 2,250 mg/day to 2,500 mg/day. In addition, the present disclosure is aimed at frictionally administering nelfinavir once to four times a day at the low dose to provide an optimal pharmaceutical use and pharmaceutical composition for relieving symptoms of COVID-19 while reducing gastrointestinal side effects, or not accompanying the gastrointestinal side effects, and not exacerbating gastrointestinal symptoms.

The pharmaceutical composition of the present disclosure may further include a carrier, an excipient, a diluent, and the like, commonly used in preparation of pharmaceutical compositions.

The "carrier" may be defined as a substance facilitating administration of a compound into a cell or tissue. The "excipient" refers to a substance added to provide an appropriate form to a drug or to increase an amount of the drug to be conveniently used. The "diluent" may be defined as a substance not only stabilizing biologically active form of the compound but also dissolving and diluting the compound.

Such a carrier, excipient, or diluent may be, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, hydroxycellulose, hydroxypropylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, crospovidone, croscarmellose sodium, water, physiological saline, buffered saline, dimethylsulfoxide (DMSO), methylhydroxybenzoate, talc, magnesium stearate, mineral oil, or the like, but is not limited thereto.

A pharmaceutical composition according to the present disclosure may be formulated as a formulation appropriate for oral administration such as a powder, a granule, a tablet, a capsule, a solution, a suspension, an emulsion, a syrup, an aerosol, or the like, and may be used as a formulation for injection administration such as subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection, or a formulation for external composition, but is not limited thereto.

When the pharmaceutical composition of the present disclosure is formulated as an orally administered composition, nelfinavir, an active ingredient, or a pharmaceutically acceptable salt thereof may be formulated as a unit dosage form containing the content (potency) of 25 mg to 1,500 mg to prevent or treat SARS-CoV-2 infection, COVID-19, for example. In detail, nelfinavir or a pharmaceutically acceptable salt thereof may be formulated as a unit dosage form containing 33 mg to 750 mg for an adult human (each having a weight of 60 kg). A formulation of the formulated unit dosage may be administered once to four times a day such that a daily dosage may be 100 mg to 1,500 mg.

On the other hand, when the pharmaceutical composition of the present disclosure is formulated as an injection, nelfinavir, an active ingredient, or a pharmaceutically acceptable salt thereof may be administered as a unit dosage formulation of 25 mg to 1,500 mg a day to prevent or treat SARS-CoV-2 infection, COVID-19, for example. In detail, nelfinavir or a pharmaceutically acceptable salt thereof may be administered as a unit dosage formulation of 33 mg to 750 mg per day for an adult human. A formulation of the formulated unit dosage may be administered once to four times a day such that a daily dosage may be 100 mg to 1,500 mg.

The dosage of the composition according to the present disclosure may be adjusted, based on a usual dose for adjusts, depending on various factors such as type of disease, severity of the disease, type and content of another ingredient contained in a pharmaceutical composition, type of formulation, a patient's age, weight, general health status, gender, and diet, administration time and route, duration of treatment, and concurrently used drugs.

Nelfinavir or a pharmaceutically acceptable salt thereof according the present disclosure may be commercially available or may be synthesized by an organic synthesis method known in the art, and a method of the preparing nelfinavir or the pharmaceutically acceptable salt thereof is not necessarily limited.

In addition, the present disclosure provides a method for preventing or treating SARS-CoV-2 infection, which includes administering nelfinavir or a pharmaceutically acceptable salt thereof as an active ingredient to a subject, requiring the same, at a low dose of 100 mg or more to 1,500 mg or less per day for an adult human. The same descriptions of the nelfinavir and the pharmaceutically acceptable salt thereof are applied to the present method.

With the low dose of nelfinavir or the pharmaceutically acceptable salt thereof, gastrointestinal side effects may be reduced or may not be accompanied, and gastrointestinal symptoms may not be exacerbated. Therefore, subjects, to which the method for preventing or treating SARS-coronavirus-2 (SARS-CoV-2) infection according to the present disclosure is applicable, may include patients who have gastrointestinal symptoms, such as abdominal pain, abdominal discomfort, diarrhea, and the like, caused by an inflammatory response intensified in gastrointestinal mucosal tissues, but the present disclosure is not limited thereto.

In the method for preventing or treating the SARS-coronavirus-2 (SARS-CoV-2) infection, the operation of administering may be conventionally performed by, in detail, systemically administering a compound, containing the compound expressed by the chemical formula 1 as an active ingredient, but the present disclosure is not limited thereto.

The systemic administration may be performed by oral or non-oral administration. The non-oral administration may be performed by injecting subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection. The systemic administration may be performed by, in detail, oral administration.

Hereinafter, the present disclosure will be described more specifically through examples. The following examples are for illustrative purposes only and are not intended to limit the scope and spirit of the present disclosure.

Example

Example 1: Evaluation of anti-coronavirus activity of nelfinavir against SARS-CoV-2 in human lung cells

Calu-3 cell line, a human lung cell line, was obtained from Korea Cell Line Bank (KCLB), and then cultured in Dulbecco's Modified Eagle's Medium (DMEM) under conditions of 37℃ and CO ₂ of 5%. Calu-3 cells were treated with various concentrations of nelfinavir mesylate (Sigma Aldrich) corresponding to 25, 12.5, 6.25, 3.125, 1.563, 0.781, 0.391, 0.195, 0.098, 0.049, 0.024, and 0.012 μM. After two hours from the treatment, the Calu-3 cells were infected with SARS-CoV-2 (NCCP43326) at multiplicity of infection (MOI) of 0.1.

After 24 hours of infection, a culture medium was taken, and then an expression level of SARS-CoV-2 in the culture medium was determined by the amount of RNA of the virus using a quantitative RT-PCR (qRT-PCR) method. FIG. 1 illustrates dose-response curves based on the amount of SARS-CoV-2 RNA depending on nelfinavir mesylate concentration. In FIG. 1, X-axis represents the treated drug concentration (μM) in log scale, and Y-axis represents the amount of virus expressed in the drug-treated group by percentage (%) when a virus-infected group without the drug treatment (a control group) is set to 100%. Based on the above, a value of IC ₅₀ of nelfinavir mesylate (a concentration at which the amount of corresponding virus is decreased by 50%) was calculated and listed in Table 1.

Comparative Example 1: Evaluation of anti-coronavirus activity of remdesivir against SARS-CoV-2 in human lung cells

Calu-3 cell line, a human lung cell line, was obtained from Korea Cell Line Bank (KCLB) and cultured in Dulbecco's Modified Eagle's Medium (DMEM) under conditions of 37℃ and CO ₂ of 5%. Calu-3 cells were treated with various concentrations of remdesivir (BLD Pharm) corresponding to 25, 12.5, 6.25, 3.125, 1.563, 0.781, 0.391, 0.195, 0.098, 0.049, 0.024, and 0.012 μM. After two hours from the treatment, the Calu-3 cells were infected with SARS-CoV-2 (NCCP43326) at multiplicity of infection (MOI) of 0.1.

After 24 hours of infection, a culture medium was taken, and then an expression level of SARS-CoV-2 in the culture medium was determined by the amount of RNA of the virus using a quantitative RT-PCR (qRT-PCR) method. FIG. 1 illustrates dose-response curves based on the amount of SARS-CoV-2 RNA depending on remdesivir concentration. In FIG. 1, X-axis represents the treated drug concentration (μM) in log scale, and Y-axis represents the amount of virus expressed in the drug-treated group by percentage (%) when a virus-infected group without the drug treatment (a control group) is 100%. Based on the dose-response curves of FIG. 1, a value of IC ₅₀ of remdesivir (a concentration at which the amount of corresponding virus is decreased by 50%) was calculated and listed in Table 1.

Comparative Example 2: Evaluation of anti-coronavirus activity of hydroxychloroquine against SARS-CoV-2 in human lung cells

Calu-3 cell line, a human lung cell line, was obtained from Korea Cell Line Bank (KCLB) and cultured in Dulbecco's Modified Eagle's Medium (DMEM) under conditions of 37℃ and CO ₂ of 5%. Calu-3 cells were treated with various concentrations of hydroxychloroquine (Sigma Aldrich) corresponding to 100, 50, 25, 12.5, 6.25, 3.125, 1.563, 0.781 and 0.391 μM. After two hours from the treatment, the Calu-3 cells were infected with SARS-CoV-2 (NCCP43326) at multiplicity of infection (MOI) of 0.1.

After 24 hours of infection, a culture medium was taken, and then an expression level of SARS-CoV-2 in the culture medium was determined by the amount of RNA of the virus using a quantitative RT-PCR (qRT-PCR) method. FIG. 1 illustrates dose-response curves based on the amount of SARS-CoV-2 RNA depending on hydroxychloroquine concentration. In FIG. 1, X-axis represents the treated drug concentration (μM) in log scale, and Y-axis represents the amount of virus expressed in the drug-treated group by percentage (%) when a virus-infected group without the drug treatment (a control group) is 100%. Based on the above, a value of IC ₅₀ of hydroxychloroquine (a concentration at which the amount of corresponding virus is decreased by 50%) was calculated and listed in Table 1.

IC ₅₀ of Compound against SARS-CoV-2

		Present Example 1	Comparative Example 1	Comparative Example 2
Antiviral Effect	IC ₅₀* (μM)	0.01	0.03	> 50
Antiviral Effect	IC ₅₀** (ng/mL)	5.7	18.1	> 16,795

*Method for measuring IC ₅₀: a drug concentration, at which the amount of virus is decreased by 50%, was calculated as IC ₅₀ from a dose-response curve, using GraphPad Prism software

**Method for calculating IC ₅₀: IC ₅₀ (ng/mL) was calculated through conversion because nelfinavir (molecular weight: 567.8) corresponded to 1 μM = 567.8 ng/mL, remdesivir (molecular weight: 602.6) corresponds to 1 μM = 602.6 ng/mL, and hydroxychloroquine (molecular weight: 335.9) corresponded to 1 μM = 335.9 ng/mL

Remdesivir and hydroxychloroquine, used in the comparative examples, are drugs which have been most extensively in the worldwide clinical trials to discover SARS-CoV-2-related therapeutic agents. Remdesivir has been reported to be effective in reducing a period of treatment for patients who were hospitalized with COVID-19 by about four days, while the effectiveness of hydroxychloroquine has been reported to be insignificant in the clinical trials.

As can be seen from Table 1 and FIG. 1, nelfinavir used in Example 1 of the present disclosure exhibited IC ₅₀ of 0.01 μM significantly effective against SARS-CoV-2, and remdesivir used in Comparative Example 1 exhibited IC ₅₀ of 0.03 μM and hydroxychloroquine used in Comparative Example 2 exhibited IC ₅₀ of 50 μM or more and, accordingly, nelfinavir of the present disclosure had the most excellent antiviral effects against SARS-CoV-2.

However, in the case of nelfinavir, cells started to be gradually detached at concentration of 10 μM or more in a virus-infected experimental system. As compared with other common drugs which are not substantially problematic even at higher concentration than that of 10 μM, nelfinavir may be considered to be a relatively cytotoxic drug.

As described in the Background Art, in the existing studies, the antiviral effects were evaluated using cell lines in which proteins such as ACE2 and TMPRSS2, the main routes of human cell infection of SARS-CoV-2, were rarely expressed or artificially overexpressed, and thus, effectiveness and low-dosage use of nelfinavir were not found. Meanwhile, in the present disclosure, improved treatment efficacy and decreased side effects at a low dose of nelfinavir were confirmed using a cell line in which the human cell infection route factors were autonomously expressed.

Based on the above test results, pharmacokinetic factors were reviewed through reference documents to examine clinical treatment efficacy. According to a document material (Viracept tablet manual, AGOURON PHARMACEUTICALS, INC.) in which the drug concentrations in blood plasma were measured after orally administering nelfinavir, maximum plasma concentration Cmax was 4,000 ng/mL and AUC was 52,800 ng·hr/mL when 1,250 mg of nelfinavir was administered twice a day. If pharmacokinetic factors, which may be formed when nelfinavir is administered at a low dose, are calculated based on the above, maximum plasma concentration Cmax is predicted to be 160 ng/mL and AUC is predicted to be 2,112 ng·hr/mL when 50 mg of nelfinavir is administered twice a day (100 mg/day), and maximum plasma concentration Cmax is predicted to be 2,400 ng/mL and AUC is predicted to be 31,680 ng·hr/mL when 750 mg of nelfinavir is administered twice a day (1,500 mg/day).

According to the Nelfinavir PK Literature (Ther Drug Monit, Vol. 23, No. 4, 2001), the drug concentration in blood plasma immediately before the next taking of nelfinavir after taking the nelfinavir, that is, the minimum plasma concentration Ctrough is quite high. Ctrough was observed to be 1,980 ng/mL when 1,250 mg of nelfinavir was taken twice daily and was observed to be 2,380 ng/mL when 750 mg of nelfinavir was taken three times daily.

According to the literature material in which plasma concentration was measured after administering remdesivir (Summary on compassionate use, Remdesivir Gilead, EMA/178637/2020-Rev.1), maximum plasma concentration Cmax was 5,440 ng/mL and AUC was 2,920 ng·hr/mL when 200mg of remdesivir was administered by intravenous infusion. For the reference, remdesivir can be administered only by intravenous infusion. In addition, according to the literature material (Rheumatol Ther, 2015, 2; 183-195) in which plasma concentration was measured after administering hydroxychloroquine, maximum plasma concentration Cmax was 34.3 ng/mL and AUC was 559 ng·hr/mL when 200 mg of hydroxychloroquine was orally administered.

When the maximum plasma concentration Cmax of each of the above drugs is divided by IC ₅₀ of each of the drugs, a low dose of nelfinavir is calculated to be 28 to 421, remdesivir is calculated to be 301, and hydroxychloroquine is calculated to be 0.004, as illustrated in Table 2. In addition, when an AUC value of each of the drugs, referring to a total amount of drugs present in blood plasma, is divided by IC ₅₀ of each of the drugs, a low dose of nelfinavir is calculated to be 371 to 5,558, remdesivir is calculated to be 161, and hydroxychloroquine is calculated to be 0.07, as illustrated in Table 2. From the viewpoint of pharmacokinetic concept, it is expected that the greater a value obtained by dividing Cmax by IC ₅₀ and a value obtained by dividing AUC by IC ₅₀, the higher a treatment efficacy. Therefore, a low dose of nelfinavir and remdesivir are expected to have effective treatment efficacies, and hydroxychloroquine is not expected to have treatment effectiveness.

Furthermore, nelfinavir is an oral formulation and maintained at high AUC because it is exhibits excellent in-vivo duration, whereas remdesivir requires drip injection through a vein for 30 minutes to 2 hours and has low AUC because fast drug elimination occurs after the injection is finished. Therefore, considering AUC, the most relevant pharmacokinetic factor with expected treatment effectiveness, the low dose of nelfinavir according to the present disclosure has a remarkably higher value, obtained by dividing the AUC by IC ₅₀, than remdesivir. As a result, excellent clinical efficacy of the low dose of nelfinavir is expected.

Moreover, in terms of uses, remdesivir is limited to inpatients and patients with severe symptoms because intravenous infusion should be continuously performed through a vein, whereas nelfinavir may be advantageously applied to a treatment for mild to moderate COVID-19 patients including outpatients as well as severe COVID-19 patients.

	Nelfinavir		Remdesivir	Hydroxychloroquine
Route of Administration	Oral		Intravenous Infusion (Drip Injection)	Oral
Dose
	100 mg/day (50mg bid)	1,500 mg/day (750mg bid)	200 mg	400 mg
Cmax (ng/mL)	160 ^a)	2,400 ^a)	5,440	68.6
AUC (ng·hr/mL)	2,112 ^a)	31,680 ^a)	2,920	1,118
Cmax(ng/mL)/IC ₅₀(ng/mL)	28	421	301	0.004 ^b)
AUC / IC ₅₀	371	5,558	161	0.07 ^b)

a) an expected value was calculated when 50 mg or 750 mg of nelfinavir is taken twice a day, based on the PK Literature material (Viracept tablet instruction manual, AGOURON PHARMACEUTICALS, INC)

b) Cmax was calculated under the assumption that an IC ₅₀ value of hydroxychloroquine is 50 μM (an actual IC ₅₀ value was observed to be higher than 50 μM)

Example 2: Evaluation of anti-coronavirus activity of nelfinavir against SARS-CoV-2 in Golden Syrian hamsters

To perform an in-vivo test, Golden Syrian hamsters (JANVIRER LABS) were infected intranasally with 200 μL of 10 ⁴ TCID ₅₀/mL of SARS-CoV-2 (NCCP43326).

Nelfinavir mesylate (TCI Co., Ltd) was orally administered twice daily at doses of 100 mg/kg/day and 200 mg/kg/day (corresponding to 85.5 mg/kg/day or 171 mg/kg/day of nelfinavir as an active ingredient) for a week to the SARS-CoV-2 infected hamsters. In the case of a control group, a vehicle (1% of Tween80 solution) was administered without nelfinavir under the same conditions.

After three days (3 dpi, days post infection), five days (5 dpi), and seven days (7 dpi), a hamsters was sacrificed to obtain a lung tissue. The amounts of SARS-CoV-2 remaining in the lung tissue were confirmed as the amounts of expression of an envelope gene (E gene), a nucleocapsid protein gene (N gene), and an RNA-dependent RNA polymerase gene (RdRP gene) of a virus using quantitative RT-PCR (qRT-PCR, Allplex ^TM 2019-nCoV assay kit, Seegene Inc.). Based on this, under the assumption that a viral load of a control group infected with a virus is 100%, a viral load (%) expressed in each treatment group was derived, and then a value obtained by subtracting a corresponding viral load (%) from 100 was calculated as a SAS-coronavirus-2 inhibition rate (%) of nelfinavir and listed in Table 3 (85.5 mg/kg/day) and Table 4 (171 mg/kg/day).

Nelfinavir 85.5 mg/kg/day	Inhibition Rate (%)
Nelfinavir 85.5 mg/kg/day	E gene	N gene	RdRp gene
3 dpi	99.789 %	99.999 %	99.874 %
5 dpi	99.926 %	99.998 %	99.955 %
7 dpi	99.984 %	99.997 %	99.868 %

Nelfinavir 171 mg/kg/day	Inhibition Rate (%)
Nelfinavir 171 mg/kg/day	E gene	N gene	RdRp gene
3 dpi	99.966 %	99.996 %	99.999 %
5 dpi	99.995 %	99.998 %	99.974 %
7 dpi	99.994 %	99.997 %	99.335 %

As illustrated in Tables 3 and 4, in Example 2 of the present disclosure, nelfinavir had an inhibition rate of at up to 99.99% against SARS-CoV-2, and exhibited excellent antiviral activity.

To convert the dose of nelfinavir administered to the hamsters (85.5 mg/kg/day or 171 mg/kg/day) into a human equivalent dose (HED), when each value is divided by 7.4 according to the draft guideline of FDA, it is converted into the same values as 11.55 mg/kg/day and 23.11 mg/kg/day in humans, respectively. Since the values correspond to 693 mg/day and 1,387 mg/day of nelfinavir for a 60kg adult human respectively, it was confirmed well, in an actual in-vivo test, that nelfinavir exhibited excellent anti-SARS-CoV-2 activity even at a significantly lower dose than an usual dose (2,500 mg or 2,250 mg/day) when nelfinavir is used as an anti-AIDS drug. In addition, it was confirmed that a low dose of nelfinavir which was orally administered efficiently inhibited the viral expression in the lung tissue (~ 99.99% inhibition rate), so that nelfinavir may be orally well absorbed and then effectively delivered to the infected tissues to have excellent efficacy.

Accordingly, it is expected that administration of a low dose of nelfinavir will exhibit excellent anti-SARS-CoV-2 effects while significantly reducing a risk of side effects of nelfinavir, and thus, will be advantageously used in treatment of COVID-19.

While example embodiments have been illustrated and described above, it will be apparent to those skilled in the art that modifications and variations could be made without departing from the scope of the present disclosure as defined by the appended claims.

Claims

A pharmaceutical composition for preventing or treating SARS-CoV-2 infection, containing nelfinavir expressed by the following chemical formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

[Chemical Formula 1]
The pharmaceutical composition of claim 1, wherein the pharmaceutical composition reduces gastrointestinal side effects or does not accompany the gastrointestinal side effects, and does not exacerbate gastrointestinal symptoms.
The pharmaceutical composition of claim 1, wherein the SARS-CoV-2 infection accompanies at least symptom caused by SARS-CoV-2, selected from the group consisting of fever, redness, loss of at least one of taste and smell, phlegm, coughing, sore throat, headache, diarrhea, abdominal pain, abdominal discomfort, hemoptysis, nausea, dyspnea, upper respiratory tract infection, lower respiratory tract infection, respiratory infection, pneumonia, nasal stuffiness, body aches, muscular pain, joint pain, chest pain, chest tightness, fatigue, chill, vertigo, lymphopenia, hemocoagulation, dysphrasia, dyscinesia, mental aberration, anxiety, depression, somnipathy, stroke, delirium, brain injury, nerve injury, liver injury, kidney injury, skin rash, skin discoloration, acute myocardial infarction, myocarditis, cardiac insufficiency, conjunctivitis, eye infection, kidney infection, liver infection, skin infection, nervous system infection, and cardiovascular infection, or is asymptomatic.
The pharmaceutical composition of claim 1, wherein the SARS-CoV-2 infection accompanies a gastrointestinal symptom, comprising at least one of diarrhea, abdominal pain, and abdominal discomfort, caused by SARS-CoV-2.
The pharmaceutical composition of claim 1, wherein nelfinavir or pharmaceutically acceptable salt thereof are not concomitantly administered with HIV protease inhibitor, a reverse transcriptase inhibitor, or a combination thereof.
The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is an orally administered composition of a formulation selected from the group consisting of a tablet, a capsule, a powder, and a liquid.
The pharmaceutical composition of claim 4, wherein the orally administered composition has a dosing regimen in which the composition is administered once a day or is divided into two to four parts to be administered twice or four times a day.
The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated as an injection composition for intramuscular administration, subcutaneous administration, or intravenous administration.
A use of nelfinavir, expressed by the following chemical formula 1, or a pharmaceutically acceptable salt thereof for preventing or treating SARS-CoV-2 infection using the nelfinavir or the pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

[Chemical Formula 1]
The use of nelfinavir of claim 9, wherein the low dose of nelfinavir or the pharmaceutically acceptable salt thereof reduces gastrointestinal side effects or does not accompany the gastrointestinal side effects, and does not exacerbate gastrointestinal symptoms.
A use of nelfinavir, expressed by the following chemical formula 1, or a pharmaceutically acceptable salt thereof for preparing a medicament for preventing or treating SARS-CoV-2 infection using the nelfinavir or the pharmaceutically acceptable salt thereof as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

[Chemical Formula 1]
The use of nelfinavir of claim 11, wherein the low dose of nelfinavir or the pharmaceutically acceptable salt thereof reduces gastrointestinal side effects or does not accompany the gastrointestinal side effects, and does not exacerbate gastrointestinal symptoms.
A method for preventing or treating SARS-CoV-2 infection, the method comprising:

administering nelfinavir, expressed by the following chemical formula 1, or a pharmaceutically acceptable salt thereof to a subject in need of treatment, as an active ingredient at a low dose of 100 mg/day to 1,500 mg/day for an adult human.

[Chemical Formula 1]
The method of claim 13, wherein the low dose of nelfinavir or the pharmaceutically acceptable salt thereof reduces gastrointestinal side effects or does not accompany the gastrointestinal side effects, and does not exacerbate gastrointestinal symptoms.