CN112516122A

CN112516122A - Application of suramin in preparation of drugs for preventing or treating coronavirus related diseases

Info

Publication number: CN112516122A
Application number: CN202011463396.5A
Authority: CN
Inventors: 申屠建中; 洪丽萍; 辜列; 周静
Original assignee: Hong Nanfang
Current assignee: Hong Nanfang
Priority date: 2020-04-21
Filing date: 2020-12-11
Publication date: 2021-03-19

Abstract

The invention belongs to the technical field of medicines. The present invention provides the use of suramin, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for preventing or treating a coronavirus-related disease in a subject. The invention also provides a medicament for preventing or treating coronavirus related diseases, which comprises suramin or a pharmaceutically acceptable salt thereof. The medicaments provided herein are useful for the prevention and treatment of novel coronavirus pneumonia.

Description

Application of suramin in preparation of drugs for preventing or treating coronavirus related diseases

Technical Field

The invention relates to the technical field of medicines, in particular to application of suramin in preparation of a medicine for preventing or treating coronavirus related diseases.

Background

"coronaviruses" refers to a class of enveloped viruses that cause disease in vertebrates, particularly mammals or birds, and have a single positive-stranded RNA genome. They belong to the phylogenetic group of the order of the nested viruses (Nidovirales), the family of Coronaviridae (Coronaviridae). The presence of spinous processes on the envelope of such viruses was observed under an electron microscope, and the whole viruses were like coronas, so that they were named coronaviruses. Animal or human diseases caused by coronaviruses are referred to as "coronavirus-related diseases" in the present invention. It has been found to date that 7 coronaviruses can infect humans, including HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, and SARS-CoV-2. Some of the diseases they cause are mild, resembling common flu, while others may be fatal, such as SARS, MERS and COVID-19.

COVID-19 is a pneumonia caused by a novel coronavirus which was discovered in 2019, and patients infected by the virus have typical symptoms such as fever, dry cough, dyspnea, headache, pneumonia and the like. Patients with mild symptoms may only experience a runny nose or sore throat. In severe patients, dyspnea, cytokine storm, and even eventual organ failure can occur. The national health committee on 2/8/2020 calls this disease "novel coronavirus pneumonia", and the world health organization announces its corresponding english name "COVID-19 (Corona VirusDisease 2019)" on 2/11/2020. Since the coronavirus causing the pneumonia and the coronavirus causing SARS are highly related in taxonomy, the coronavirus research group (CSG) of the International Committee for viral Classification named the virus "SARS-CoV-2". In some literature, the virus is also referred to as "2019-nCoV". To date, no vaccine or drug has been approved for the prevention or treatment of this disease.

Suramin (Suramin), also known as Bayer-205, is a polyaryl urea polysulfonate. Its hexasodium salt (suramin sodium, C)₅₁H₃₄N₆Na₆O₂₃S₆) Is white to light yellow with high water solubility and hygroscopicityAnd (3) powder. Suramin has been used for the treatment of african trypanosomiasis and onchocerciasis since 1919 and was included by the World Health Organization (WHO) in the "basic drug catalogue of adults" and the "basic drug catalogue of children under 12 years old", respectively. Scholars in the 80's of the 20 th century discovered that suramin could inhibit the activity of reverse transcriptase associated with oncovirus, and used suramin in the treatment of aids. Although suramin is discovered to be not an effective drug for treating AIDS later, suramin is unexpectedly found to block the activities of certain growth factors and enzymes closely related to tumor growth in the treatment process, so that suramin as an anti-tumor drug is widely applied to the treatment of various malignant tumors. In recent years, other therapeutic applications of suramin have been increasingly discovered, for example, as P2X receptor antagonists for the treatment of ocular proliferative diseases and autism in children. Recent findings also include that suramin sodium inhibits strains related to the enterovirus coxsackievirus type a, and is therefore useful in the treatment of hand-foot-and-mouth disease in children.

Suramin has the following structure:

suramin derivatives (including analogs thereof) have been reported in the prior art, including, for example, NF110, NF032, NF201, NF023 and NF103 disclosed in US6121320A, and NF023, NF279 and NF157 disclosed in chinese patent CN 103301462A. A number of suramin-type compounds are also disclosed in PCT application publication WO9015816A1, and reference is made specifically to the description on pages 4-5 of the specification. Processes for preparing the above suramin derivatives and other suramin derivatives are also known in the prior art, and are described, for example, in Nickel P, Haack H J, Widjajaja H, et al [ Potential filitics. suramin alloys ] [ J ]. Arzneimittel-Forschung,1986,36(8):1153.

At present, no report on the application of suramin and derivatives thereof in preventing or treating COVID-19 exists. Therefore, it is of great importance to develop a drug that can effectively prevent or treat the disease.

Disclosure of Invention

Aiming at the defects in the prior art, the technical problem to be solved by the invention is to provide an anti-coronavirus drug which can effectively prevent or treat diseases related to coronavirus.

In order to solve the technical problems, the invention provides the following technical scheme:

in one aspect, the present invention provides the use of suramin, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for preventing or treating a coronavirus-related disease in a subject.

In some embodiments, the pharmaceutically acceptable salt of suramin is suramin sodium.

In some embodiments, the coronavirus is HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV, or SARS-CoV-2.

In some embodiments, the coronavirus is the strain BetacoV/Guangdong/20SF014/2020(EPI _ ISL _ 403934).

In some embodiments, the coronavirus related disease is COVID-19.

In some embodiments, the subject is a mammal or bird; the mammal is a human.

Furthermore, in the case where suramin is disclosed as having anti-neocoronaviral activity, one skilled in the art would predict that these suramin derivatives may also have anti-neocoronaviral activity. Therefore, the application of the suramin derivatives with the activity of resisting the new coronavirus in preparing the medicines for preventing and/or treating the diseases related to the new coronavirus is also included in the protection scope of the invention.

In another aspect, the present invention provides a medicament for preventing or treating a coronavirus-related disease in a subject, said medicament comprising suramin or a pharmaceutically acceptable salt thereof.

In some embodiments, the therapeutically effective amount of suramin sodium is 15-20mg suramin sodium/kg body weight.

Further, the therapeutically effective amount of suramin sodium is 15mg suramin sodium/kg body weight.

In another aspect, the present invention provides a pharmaceutical kit for preventing or treating a coronavirus-related disease, said pharmaceutical kit comprising suramin or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention provides an inhibitor of coronavirus replication in a cell, said inhibitor comprising suramin or a pharmaceutically acceptable salt thereof.

Compared with the prior art, the invention has the following beneficial effects:

the invention discovers that suramin sodium has high inhibitory activity on the new coronavirus, has the early effect of inhibiting the new coronavirus from entering cells, and has the tendency of accelerating the conversion of viral nucleic acid to negative. Therefore, suramin or pharmaceutically acceptable salts thereof can be used for preparing medicaments for preventing or treating diseases related to the coronavirus, and theoretical basis and a new way are provided for preventing and treating the new coronavirus.

Drawings

FIG. 1 shows the cytotoxicity of different concentrations of suramin sodium on Vero-E6 cells.

FIG. 2 shows the cytotoxicity of Vero-E6 cells with different concentrations of Reidcisvir and chloroquine phosphate.

FIG. 3 is a schematic diagram of the experimental design of the in vitro cell virus replication experiment.

FIG. 4 shows the inhibitory activity of different concentrations of suramin sodium on viral replication.

FIG. 5 shows the inhibitory activity of chloroquine phosphate at various concentrations on viral replication.

Figure 6 shows the inhibitory activity of redciclovir on viral replication at different concentrations.

FIG. 7 is a graph showing the effect of viral infection on the state of cells in culture.

Detailed Description

The present invention will be further illustrated in detail with reference to the following specific examples, which are not intended to limit the present invention but are merely illustrative thereof. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.

In the examples below, in vitro cell infection experiments were performed using the specific strain BetacoV/Guangdong/20SF014/2020(EPI _ ISL _ 403934).

Terms and explanations:

in the present invention, a method of preventing or treating a coronavirus-related disease in a subject is disclosed, comprising administering to the subject a therapeutically effective amount of suramin, or a pharmaceutically acceptable salt thereof.

In the present invention, a method of preventing or treating a coronavirus-related disease in a subject is disclosed, comprising administering to the subject a therapeutically effective amount of suramin, or a pharmaceutically acceptable salt thereof, in a medicament.

As used herein, "subject" refers to an individual who has or is suspected of having a disease (e.g., COVID-19), or, in predicting risk of developing a disease, may also include healthy individuals. The term is often used interchangeably with "patient", "test subject", "treatment subject", and the like. In most embodiments of the invention, the subject is a vertebrate, preferably a mammal and/or bird, more preferably a human.

"preventing" refers to avoiding, reducing, or delaying the appearance of a particular disease or disease-related symptom in a subject, and the absence of such disease or disease-related symptom prior to administration of the relevant agent. "preventing" does not require complete prevention of the occurrence of a disease or disease-related symptoms, e.g., reducing the risk of a subject developing a particular disease or disease-related symptoms after administration of a relevant drug, or lessening the severity of later-occurring related symptoms, can be considered "preventing" the occurrence or progression of the disease. "treating" or "treatment" refers to ameliorating, reducing, ameliorating, or inhibiting (e.g., arresting the development of) a disease that a subject has exhibited or has experienced. For a particular disease, "treating" may include "curing" the disease, but in most cases it is not necessary to completely eliminate all of its symptoms, e.g., administration of a related drug results in the reduction or elimination of at least one symptom in the subject, which may be considered to be treatment of the subject.

A "therapeutically effective amount" refers to an amount sufficient to elicit the biological or medical response in a subject that is desired by a clinician and can generally be determined by one of skill in the art based on factors such as the route of administration, the weight of the subject, the age, the condition, etc. When used for prophylactic purposes, a "therapeutically effective amount" can also be considered a "prophylactically effective amount". For example, a typical daily dose may range from 0.01 μ g to 100mg of the pharmaceutically active ingredient per kg body weight of the subject.

"pharmaceutically acceptable salts" refers to inorganic or organic acid addition salts and inorganic or organic base addition salts that are substantially non-harmful to the animal or human body. Common acid addition salts are, for example, but not limited to, hydrochloride, hydrobromide, nitrate, perchlorate, phosphate, sulfate, formate, acetate, aconate, ascorbate, benzenesulfonate, benzoate, cinnamate, citrate, heptanoate, fumarate, glutamate, glycolate, lactate, maleate, malonate, mandelate, methanesulfonate, naphthalene-2-sulfonate, phthalate, salicylate, sorbate, stearate, succinate, tartrate, p-toluenesulfonate and the like. Common base addition salts are exemplified by, but not limited to, ammonium salts, alkali metal salts (e.g., sodium and potassium salts), alkaline earth metal salts (e.g., calcium and magnesium salts), organic base salts (e.g., isopropylamine salts, dicyclohexylamine salts, diethylamine salts, and the like). Such salts may be prepared by methods well known to those skilled in the art. For suramin, the preferred salts are the sodium salts, especially the hexasodium salt.

In some embodiments, pharmaceutically active ingredients such as suramin may be used alone or formulated with pharmaceutically acceptable carriers into pharmaceutical compositions for use.

"pharmaceutically acceptable carrier" refers to a solid or liquid diluent, filler, antioxidant, stabilizer, etc., which is safe for administration to the animal or human body and which is suitable for administration to humans and/or animals without undue adverse side effects, while maintaining the viability of the drug or active ingredient located therein. Depending on the route of administration, a variety of different carriers well known in the art may be employed. Common routes of administration include, for example, oral, intravenous infusion, intramuscular injection, subcutaneous injection, subdominal, rectal, sublingual, or by inhalation, transdermal, and the like. Commonly used carriers include, but are not limited to, sugars, starches, cellulose and its derivatives, maltose, gelatin, talc, calcium sulfate, vegetable oils (e.g., castor oil), synthetic oils, polyols, alginic acid, phosphate buffers, emulsifiers, isotonic saline and/or pyrogen-free water, and the like.

The pharmaceutical composition can be prepared into any clinically acceptable dosage forms, such as tablets, granules, powders, capsules, injection preparations, suppositories, eye drops, external application ointments, medicinal oils, or sprays, and the like.

The above-mentioned "pharmaceutical combination" refers to a combination of two or more pharmaceutically active ingredients. In the case of a pharmaceutical combination comprising two pharmaceutically active ingredients, it does not only comprise the co-presence of the two pharmaceutically active ingredients in the same pharmaceutical formulation (e.g. pharmaceutical composition), but also means that the two pharmaceutically active ingredients may be present in the same pharmaceutical kit as separate pharmaceutical formulations, or even that the two pharmaceutically active ingredients may be present in different pharmaceutical kits as separate pharmaceutical formulations. In fact, such administration is intended to be a "drug combination" as long as the two pharmaceutically active ingredients are capable of being present in the subject at the same time by administration. In most cases, when two pharmaceutically active ingredients are constituted into a pharmaceutical combination and administered in combination, the two pharmaceutically active ingredients exert a synergistic effect in the subject. Such synergistic effects include a combination that results in a better efficacy than either of the pharmaceutically active ingredients alone, one of the pharmaceutically active ingredients may reduce the side effects of the other, or a combination of two pharmaceutically active ingredients that results in new efficacy, e.g., for the treatment of an existing indication different from either of the pharmaceutically active ingredients. These pharmaceutically active ingredients are also optionally formulated with a pharmaceutically acceptable carrier.

In other words, a pharmaceutical combination includes not only a pharmaceutical composition form but also other forms in which different pharmaceutically active ingredients may be present in a subject.

Some aspects of the present invention are based, at least in part, on the inventors' unexpected discovery through in vitro cell experiments and clinical trials that suramin, or a pharmaceutically acceptable salt thereof, has anti-coronavirus effects in vitro and in vivo. These in vitro experiments and clinical trials are described in further detail below.

The inventor investigates the sensitivity of suramin sodium and other three drugs to the new coronavirus 2019n-CoV through in vitro cell experiments, and the method comprises the steps of detecting the number of living cells through CCK-8 and calculating CC50 (50% cytoxic concentration); EC50 (50% effect concentration) was calculated by RT-PCR to quantify the number of viruses in the cell supernatant. The test virus strain was a covv-19 virus strain isolated from the clinic and sequenced through the whole genome (see below).

EXAMPLE 1 determination of drug cytotoxicity

1.1 materials

Medicine preparation: suramin sodium (kang Zhi pharmaceutical industry, lot number: 03076-20190601), Reidesvir (ChemTown, CT2020021010), ribavirin (Guangdong lake Biochemical pharmaceutical factory), and chloroquine phosphate.

Cell: Vero-E6.

Cell culture solution: 10% FBS 1% PSMEM medium (Gibico, Cat No. 42360032).

Cell maintenance solution: 2% FBS 1% PSMEM medium (Gibico, Cat No. 42360032).

1.2 methods

Drug dilution: the drug to be tested was diluted in 96-well plates in cell culture medium in stepwise manner. Each concentration was paralleled by 3 wells with cell control wells (no drug, culture medium only). The ratio of the number of viable cells of the drug groups at different concentrations to the control of the blank cells was determined by CCK-8 reagent after 48 hours to calculate the CC50 value for each drug.

1.3 results

The CC50 value of suramin sodium to Vero-E6 cells was 306. mu.M; the CC50 value of chloroquine phosphate to Vero-E6 cells is 326 mu M; the CC50 value of Reidesciclovir on Vero-E6 cells is 40. mu.M (see FIGS. 1 and 2).

Example 2 pharmacodynamic test of four drugs such as suramin to inhibit 2019-nCoV virus

2.1 materials

The cells and cell culture medium used were as in example 1, and the cell maintenance medium was 2% FBS 1% PSMEM medium.

Virus strain: the 2019-nCoV strain isolated from CDC of Guangdong province has complete genome sequence data shown in GISAID database with the sequence number EPI _ ISL _403934 and the strain number BetacoV/Guangdong/20SF 014/2020.

The detection kit comprises: 2019-nCoV virus fluorescent quantitative RT-PCR kit (Daan gene).

2.2 methods

Vero-E6 cells were cultured in advance in 96-well plates to 2X 10 monolayers⁴A hole. The virus infectious titer was MOI0.01, 7 drug concentrations (see table 1 below) were chosen, and 3 experiments were repeated at each concentration. Referring to fig. 3, the test was performed as follows:

test # 1: incubating virus and medicine for 1 hour, infecting cells, washing virus and culture solution containing medicine after infecting for 2 hours, and adding cell maintenance solution after washing once; the results data were used to compare the effect of different drugs on virus Entry (Entry).

Test # 2: drugs were added at the same Time as the virus infected cells, and after 2 hours of infection, the virus was washed out while maintaining the drug concentration, and the antiviral effects of each drug on the whole virus (Full-Time) were compared with the result data.

Test # 3: after 2 hours of virus infection of cells, the virus was washed out, and then drugs were added to compare the inhibitory effect of different drugs on the virus after virus infection with the result data (Post-entry).

No drug (Virus Con.) was added to the Virus control group.

TABLE 1 drug concentrations used in the test (unit: μ M)

	Suramin sodium salt	Ribavirin (ribavirin)	Chloroquine phosphate	Ruidexiwei (Ridexil)
					1	150	600	75	30
2	75	300	37.5	15
					3	37.5	150	18.75	7.5
4	18.75	75	9.375	3.75
					5	9.375	37.5	4.6875	1.875
6	4.6875	18.75	2.34375	0.9375
					7	2.34375	9.375	1.171875	0.46875

2.3 results

Cytopathic effect (CPE) was first observed continuously every 24 hours under a P3 laboratory microscope. Observations showed that after 48 hours of infection, the virus control group developed observable cytopathic effects as a sparse number of live cells under the mirror, with the remaining cells shrinking in appearance, rounding off to fragmentation (see fig. 7). Whereas virus-free infected cells grew adherent as a complete monolayer. However, when different concentrations of suramin sodium (see the concentration mark in fig. 7) were added to the virus-infected cell culture fluid, it was observed that the cell growth gradually approached the virus-free state with increasing concentrations of suramin sodium, indicating that suramin sodium can inhibit the toxic effect of the virus on cell growth.

Meanwhile, at 48 hours after infection, supernatants of all experimental groups are collected, the relative quantity of 2019n-CoV virus is calculated through fluorescent quantitative RT-PCR reaction, and the mean value and standard deviation of three experimental repeated wells are calculated at each concentration. EC50 is the half effective concentration of the drug, i.e., the effective concentration of the drug at which the viral load is suppressed to half of that of control cells.

The results show that the incubation of suramin sodium drug with the new coronavirus for 1 hour can significantly inhibit the subsequent invasion of the virus (EC50 ═ 4.19 μ M); the same inhibition was observed on the virus both at the time of infection with the neocoronaviruse (EC50 ═ 8.29 μ M) and 2 hours after infection (EC50 ═ 8.09 μ M), with no significant difference suggesting that viral adsorption and entry into the cells may be a longer process and the effect of suramin sodium may be sustained. The SI of suramin sodium inhibition was between 36.91 and 72.9 (FIG. 4).

No inhibitory effect of ribavirin on the new coronavirus was observed in any of the three experiments under drug action conditions at a maximum concentration of 600. mu.M.

The subsequent invasion of the virus can be obviously inhibited by incubating chloroquine phosphate with the new coronavirus for 1 hour (EC50 is 2.157 mu M); the simultaneous infection with the new coronaviruse (EC50 ═ 0.952 μ M) and the 2 hours after infection with the drug also showed inhibition of the virus (EC50 ═ 1.709 μ M) with higher SI, between 151 and 190.75 (fig. 5).

Resiscivir was inhibited by incubation with neocoronavirus for 1 hour (EC50 ═ 0.35 μ M), but was less effective than viral infection with concomitant drug addition and maintenance (EC50 ═ 0.145 μ M), suggesting that the drug may be more inhibitory to viral replication in the virus cells (fig. 6).

The above results are shown in table 2.

TABLE 2 suramin sodium and other drugs sensitive to the New coronavirus 2019n-CoV

The results of the in vitro cell tests of examples 1 and 2 above demonstrate that suramin sodium has a high inhibitory activity against the novel coronaviruses, with EC50 between 4.196 and 8.291 μ M and SI between 36.91 and 72.9 under the experimental conditions of the present invention. Wherein, the incubation with the new coronavirus for 1 hour can obviously inhibit the invasion of the virus, and the earlier the medicine is added, the greater the inhibition effect is. The literature reports that the mechanism of inhibition of the enterovirus 71 (which is a single-strand positive-strand RNA virus like the new coronaviruses) by suramin sodium is to neutralize the virus by binding with a 5-fold symmetrical vertex region positively charged by the capsid protein of the EV-A71 virus, thereby preventing the virus from entering host cells and inhibiting the virus from replicating. From the comparison of the results of the three experimental designs in the present study, it is suggested that suramin sodium has an early effect of inhibiting the entry of new coronavirus into cells, but the specific mechanism of action needs to be further studied.

The invention also synchronously compares the antiviral drugs chloroquine phosphate and ribavirin recommended by the national Weijian Commission on diagnosis and treatment plan of New coronary pneumonia, and the inhibitory activity of the Redexilvir, a new medicine in the research of the United states Gilead company, on the new coronary viruses. Wherein the inhibitory activities of chloroquine phosphate and Reidcisvir are close to those reported in the public (Table 2), which indicates the reliability of the activity assay system of the present invention. However, in this experiment, no inhibition was observed up to 600. mu.M in ribavirin, which is different from the data reported in Table 3 for unknown reasons. In summary, the present invention demonstrates that suramin sodium has inhibitory effects on new coronaviruses and suggests that the earlier the administration, the greater the potential for clinical benefit.

Table 3 reports on the anti-COVID-19 virus susceptibility of different drugs

Name of drug	EC50(μM)	CC50(μM)	Inhibition index SI	Data source
					Redcisvir Remdesivir	0.77	100	129.87	Wuhan virus institute
Chloroquine phosphate Chloroquine	1.13	100	88.5	Wuhan virus institute
					Nitazoxanide	2.12	35.53	16.76	Wuhan virus institute
Favipiravir	61.88	400	6.46	Wuhan virus institute
					Nafamostat	22.5	100	4.44	Wuhan virus institute
Penciclovir Penciclovir	95.96	400	4.17	Wuhan virus institute
					Ribavirin	109.5	400	3.65	Wuhan virus institute

EXAMPLE 3 clinical study of efficacy and safety of suramin sodium for injection

This example employs a multi-center, open, single arm study.

The subjects to be enrolled must meet all of the following criteria simultaneously:

(1) the patient is 18-75 years old, unlimited in nature, and voluntarily signs an informed consent;

(2) suspected cases are judged according to comprehensive analysis of comprehensive epidemiological history and clinical manifestations of pneumonia diagnosis and treatment plan (trial sixth edition) infected by novel coronavirus;

(3) detecting the positive of the novel coronavirus nucleic acid by real-time fluorescence RT-PCR; or viral gene sequencing, with high homology to known novel coronaviruses;

(4) patients with pneumonitis confirmed by chest imaging;

(5) the time interval between symptom onset and randomized inclusion was within 10 days (including 10 days); the symptom onset mainly takes fever as a judgment basis, and cough or other related symptoms can be used for patients without fever since the onset;

(6) patients had the ability to follow the study protocol at the discretion of the investigator.

Subjects must be excluded if they meet any of the following criteria:

(1) allergic constitution, known to be allergic to sodium suramin for injection or other 2 or more drugs (including but not limited to penicillins, cephalosporins, levofloxacin, clindamycin, ribavirin, pipemidic acid, analgin, compound sulfamethoxazole, etc.) or food;

(2) patient weight <40 kg;

(3) critical illness that meets one of the following conditions: respiratory failure occurs and mechanical ventilation is required; the occurrence of shock; ICU monitoring treatment is needed when other organ failure is combined;

(4) patients with impaired renal function (estimated creatinine clearance <60mL/min (male: Cr (mL/min) ═ 140-age x body weight (kg)/72 x blood creatinine concentration (mg/dl); female: Cr (mL/min) ═ 140-age x body weight (kg)/85 x blood creatinine concentration (mg/dl));

(5) patients were tested for abnormalities in any of the following laboratory parameters (according to local laboratory reference) during the screening period:

ALT or AST levels >5 times the upper limit of normal range (ULN) or

ALT or AST levels >3 times ULN and total bilirubin levels >2 times ULN;

(6) pregnant or lactating women;

(7) the investigator determines that there are other factors that may cause the subject to be forced to terminate the study, such as other serious illnesses, serious laboratory abnormalities, and other factors that affect the subject's safety or experimental data and blood sample collection.

A total of 8 patients were enrolled using the above criteria, of which 5 males (62.5%) and 3 females (37.5%) aged 45.4 + -10.0 years, 163.4 + -5.6 cm in height and 62.4 + -12.1 kg in weight, all nationalities being Han nationality. The clinical classification is the common type.

Adopts suramin sodium for injection (Jiangsu Jinsili), the dosage form is freeze-dried powder injection, and the specification is as follows: 300 mg.

Patients were admitted to the group as soon as possible after screening for suramin sodium treatment by injection.

The usage and dosage are as follows:

dosage: 15mg/kg

(1) Preparing the medicine: dissolving the medicinal preparation (300 mg/bottle, lyophilized powder for injection) into 10% isotonic solution with water for injection, mixing with 100mL physiological saline, preparing before use, and storing at room temperature in dark place;

(2) the use method comprises the following steps: intravenous drip, the infusion time is not less than 1 hour;

(3) the course of treatment is as follows: 1 dose on

day

1 and 1 dose again on day 5.

A total of 8 patients received suramin sodium for injection, of which 3 received 2 times suramin sodium for injection (D1, D5 dosing) and 5 received 1 time suramin sodium for injection (D1 dosing): in 1 case, D5 was rejected, in 1 case, adverse events with underlying cardiovascular disease occurred after D1 administration, investigators assessed no further administration of the second dose, and in 3 cases, virus nucleic acid was discharged from hospital after completion of the 1 st dose. Here, D1 and D5 refer to the first day and the fifth day, respectively.

By 3 months and 14 days in 2020, the rate of conversion of virus nucleic acid to negative of patients is 100 percent, and the discharge cure rate is 100 percent.

Statistics show that the median time from onset to discharge of 8 patients is 40(27-54) days; median time from start of antiviral treatment to discharge was 26.5(15-48) days; the median time from the start of antiviral treatment to viral negative conversion (RT-PCR) was 25(13-47) days; the time from the group administration of the suramin sodium for injection to the discharge median is 7.5(2-11) days; sulamine sodium for injection was administered to the group until the median time for viral nucleic acid to turn negative was 4.5(0-10) days. Antiviral drugs were combined during hospitalization in 8 patients: lopinavir ritonavir tablets, baroxavir disoproxil tablets, abidol tablets, chloroquine phosphate tablets, ribavirin injection and recombinant interferon a-2b spray.

The results of the above clinical studies indicate that suramin sodium shows a certain activity against COVID19 virus, and has a tendency to accelerate the turning of viral nucleic acid to negative, including patients who have not turned negative in the past by other antiviral treatments, but still needs to increase enough instances and stringent random controls to reach a conclusion of more practical clinical significance.

Finally, it should be noted that the above-mentioned contents are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, and that the simple modifications or equivalent substitutions of the technical solutions of the present invention by those of ordinary skill in the art can be made without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. Use of suramin or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for preventing or treating a coronavirus-related disease in a subject.

2. The use according to claim 1, wherein the pharmaceutically acceptable salt of suramin is suramin sodium.

3. The use of claim 1 or 2, wherein the coronavirus is HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV, MERS-CoV or SARS-CoV-2.

4. The use according to claim 1 or 2, wherein the coronavirus related disease is COVID-19.

5. Use according to claim 1 or 2, wherein the subject is a mammal or bird; the mammal is a human.

6. A medicament for preventing or treating a coronavirus related disease in a subject, said medicament comprising suramin or a pharmaceutically acceptable salt thereof.

7. The medicament of claim 6, wherein the pharmaceutically acceptable salt of suramin is suramin sodium.

8. The medicament of claim 7, wherein the therapeutically effective amount of suramin sodium is 15-20mg of suramin sodium per kg of body weight.

9. A pharmaceutical kit for preventing or treating a disease associated with a coronavirus, wherein the pharmaceutical kit comprises suramin or a pharmaceutically acceptable salt thereof.

10. An inhibitor of coronavirus replication in a cell, said inhibitor comprising suramin or a pharmaceutically acceptable salt thereof.