阅读说明：本技术 苯甲酸酯类化合物在治疗SARS-CoV-2感染方面的应用 (Application of benzoate compound in treating SARS-CoV-2 infection ) 是由 钟武 曹瑞源 肖庚富 胡志红 王曼丽 张磊砢 李薇 李月香 赵磊 樊士勇 李松 于 2020-07-15 设计创作，主要内容包括：本发明涉及式I所示苯甲酸酯类化合物、其几何异构体、其药学上可接受的盐和/或其溶剂化物或其水合物,及含上述化合物的药物组合物,用于治疗SARS-CoV-2感染方面的用途,<Image he="406" wi="700" file="DDA0002586005400000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The invention relates to benzoate compounds shown in formula I, geometric isomers, pharmaceutically acceptable salts and/or solvates or hydrates thereof, and a pharmaceutical composition containing the compounds, which are used for treating SARS-CoV-2 infection,)

1. The use of a compound of formula I, its geometric isomers, its pharmaceutically acceptable salts and/or solvates and/or hydrates thereof in the preparation of a medicament for the prevention and/or treatment of diseases or infections caused by SARS-CoV-2, including but not limited to respiratory diseases (e.g. simple infections such as fever, cough and angina, pneumonia, acute or severe acute respiratory infections, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis and septic shock, etc.),

2. use of a pharmaceutical composition for the manufacture of a medicament for the prevention and/or treatment of a disease or infection caused by SARS-CoV-2, such as a respiratory disease (e.g. simple infections such as fever, cough and angina and the like, pneumonia, acute or severe acute respiratory infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis and septic shock and the like), wherein the pharmaceutical composition comprises a compound represented by formula I, a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof,

preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant, and specifically, the pharmaceutical composition is a solid preparation, an injection, an external preparation, a spray, a liquid preparation, or a compound preparation.

3. A compound of formula I,

The use of a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof for the preparation of a medicament as a SARS-CoV-2 inhibitor.

4. The application of the pharmaceutical composition in preparing the medicine used as SARS-CoV-2 inhibitor,

wherein the pharmaceutical composition comprises a compound shown as a formula I, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof,

preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant, and specifically, the pharmaceutical composition is a solid preparation, an injection, an external preparation, a spray, a liquid preparation, or a compound preparation.

5. A compound of formula I,

The use of a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof for the manufacture of a medicament for inhibiting the replication or propagation of SARS-CoV-2 in a cell (e.g. a mammalian cell).

6. Use of a pharmaceutical composition in the manufacture of a medicament for inhibiting the replication or propagation of SARS-CoV-2 in a cell (e.g., a mammalian cell),

wherein the pharmaceutical composition comprises a compound shown as a formula I, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof,

preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant, and specifically, the pharmaceutical composition is a solid preparation, an injection, an external preparation, a spray, a liquid preparation, or a compound preparation.

7. The use of claim 1 or 2, wherein the SARS-CoV-2 caused disease is COVID-19.

8. The use of claim 5 or 6, wherein the mammal comprises a bovine, equine, ovine, porcine, canine, feline, rodent, primate, such as a human, feline or porcine.

9. Use according to any one of claims 1 to 8, wherein the pharmaceutically acceptable salt of the compound of formula I is an inorganic or organic acid salt of the compound, such as hydrochloride, hydroamidate, hydrorosate, nitrate, sulfate, bisulfate, phosphate, biphosphate, acetate, propionate, butyrate, oxalate, pivalate, adipate, alginate, lactate, citrate, tartrate, succinate, maleate, fumarate, picrate, aspartate, gluconate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate and the like.

10. The use of claim 9, wherein the pharmaceutically acceptable salt of the compound of formula I is a mesylate salt of the compound.

Technical Field

The invention relates to a benzoate compound shown in the following formula I, a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof or a hydrate thereof, and a pharmaceutical composition containing the compound, and application of the benzoate compound in the aspect of treating SARS-CoV-2 infection.

Background

Benzoate compounds of formula I, also known as Nafamostat (Nafamostat), with chemical name 6-amidino-2-naphthyl-4-guanidinobenzoate, were originally developed by the company Torii Pharma, japan, in 1986, and indicated for: acute pancreatitis, disseminated intravascular coagulation DIC, and anticoagulant.

Nafamostat is a fast acting proteolytic inhibitor that prevents the hydrolysis of fibrinogen to fibrin during hemodialysis by competitively inhibiting a variety of serine proteases, including thrombin. It can improve acute pancreatitis and prevent blood clot formation in vivoThe external circulation and the external body have anti-inflammatory effect. Nafamostat has neuroprotective effect in cerebral ischemia injury caused by antithrombin activity. In addition, nafamostat has also been reported to have multiple pharmacodynamic activities: antiproliferative effects, inhibition of cell adhesion and invasion, and increased susceptibility to anoikis were observed in nafamostat-treated cancer cells; nafamostat is effective in inhibiting interferon-gamma ray (IFN- γ) -induced upregulation of messenger rna (mrna) and PD-L1 at the protein level in human lung cancer cells (HLC-1); nafamostat plays a therapeutic role in sensitizing airway inflammation, not only because they have inhibitory effects early in mast cell activation, but also because they have immunomodulatory functions late in sensitizing inflammation. In terms of antiviral activity, nafamostat is a membrane fusion inhibitor identified as the MERS coronavirus S protein, superior to several other clinically approved serine protease inhibitors. Nafamostat can also block MERS-CoV infection in vitro, and in vitro experiments show that IC of nafamostat₅₀0.1 μ M, 10 times higher than camostat.

2019 the novel coronavirus (2019-nCoV) is a new strain of coronavirus that has not previously been found in humans. On day 11 of 2020, month 2, the international committee for viral classification (ICTV) announced that the formal classification of the 2019 novel coronavirus (2019-nCoV) was named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). On the same day, the World Health Organization (WHO) announced that the formal name of the disease caused by this virus was COVID-19. The symptoms of SARS-CoV-2 infection are mainly pneumonia, and can be classified into simple infection, mild pneumonia, severe pneumonia, acute respiratory distress syndrome, sepsis, septic shock, etc. according to the severity of the disease. Patients with simple infections may have non-specific symptoms such as fever, cough, sore throat, nasal congestion, weakness, headache, muscle aches or discomfort, and the elderly and immunosuppressors may have atypical symptoms. Patients with mild pneumonia mainly have cough, dyspnea and tachypnea. Severe pneumonia can be seen in adolescents, adults or children, and the main symptoms are increased respiratory frequency, severe respiratory failure or dyspnea, cyanosis, lethargy, unconsciousness or convulsion, air suction and the like. The lung image of acute respiratory distress syndrome is a bilateral vitreography, but cannot be completely explained by effusion, lobular effusion or atelectasis or a lung mass image, and pulmonary edema is the main symptom. Septic patients often have fatal organ dysfunction, and septic shock is the most critical patient and has a high probability of death.

At present, aiming at SARS-CoV-2 infection, the clinical treatment is mainly supported, and no anti-virus medicine is available.

Disclosure of Invention

The invention aims to discover a compound with antiviral activity on SARS-CoV-2, thereby developing a remedy for SARS-CoV-2 infection. The invention discovers that the compound shown in the formula I has the function of inhibiting the reproduction of SARS-CoV-2 through creative research, and has good potential treatment effect on the aspect of treating diseases caused by SARS-CoV-2.

The invention provides a compound shown as a formula I, a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof or a hydrate thereof:

according to the invention, the pharmaceutically acceptable salts of the compounds of formula I of the present invention include inorganic or organic acid salts and inorganic or organic base salts thereof, and the present invention relates to all forms of the above salts. Including but not limited to: sodium, potassium, calcium, lithium, meglumine, hydrochloride, hydrabamate, hydrarginate, nitrate, sulfate, bisulfate, phosphate, biphosphate, acetate, propionate, butyrate, oxalate, pivalate, adipate, alginate, lactate, citrate, tartrate, succinate, maleate, fumarate, picrate, aspartate, gluconate, benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, pamoate and the like.

In certain embodiments, the pharmaceutically acceptable salt of the compound of formula I is nafamostat mesylate.

According to the invention, the compound of formula I can inhibit the replication of SARS-CoV-2 virus on cells and reduce the nucleic acid load of SARS-CoV-2 virus in cell culture.

After a great deal of creative invention research, the inventors of the present invention found out a new action characteristic of the compound of formula I in cells: the compound of formula I can reduce the viral nucleic acid load level of SARS-CoV-2 virus infected cells at micromolar concentrations.

The invention also relates to the application of the compound shown in the formula I, the geometric isomer thereof, the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in the preparation of medicines for preventing and/or treating diseases or infections caused by SARS-CoV-2 (including but not limited to respiratory diseases (such as simple infections, such as fever, cough, angina and the like, pneumonia, acute or severe acute respiratory tract infection, hypoxic respiratory failure, acute respiratory distress syndrome, sepsis, septic shock and the like)),

the invention also relates to application of the compound shown in the formula I, geometric isomers, pharmaceutically acceptable salts and/or solvates and/or hydrates thereof in preparing medicines used as SARS-CoV-2 inhibitors.

The invention also relates to application of the compound shown in the formula I, geometric isomers thereof, pharmaceutically acceptable salts thereof and/or solvates thereof and/or hydrates thereof in preparing a medicament for inhibiting the replication or reproduction of SARS-CoV-2 in cells (such as mammalian cells).

The invention also relates to a pharmaceutical composition which comprises the compound shown in the formula I, a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof,

preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant, and specifically, the pharmaceutical composition is a solid preparation, an injection, an external preparation, a spray, a liquid preparation, or a compound preparation.

The invention also relates to application of the pharmaceutical composition containing the compound shown in the formula I, the geometric isomer thereof, the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof or the compound shown in the formula I, the geometric isomer thereof, the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing medicines for preventing and/or treating respiratory system diseases (simple infections such as fever, cough, angina and the like, pneumonia, acute or severe acute respiratory tract infection, hypoxic respiratory failure, acute respiratory distress syndrome, sepsis, septic shock and the like) but not limited by respiratory system diseases.

The invention also relates to the use of the pharmaceutical composition in the preparation of a medicament for preventing and/or treating diseases or infections caused by SARS-CoV-2, such as respiratory diseases (e.g. simple infections such as fever, cough, and angina, pneumonia, acute or severe acute respiratory infections, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock, etc.).

The invention also relates to the application of the pharmaceutical composition in preparing medicines used as SARS-CoV-2 inhibitors.

The invention also relates to the use of the pharmaceutical composition in the preparation of a medicament for inhibiting the replication or propagation of SARS-CoV-2 in a cell (e.g., a mammalian cell).

The present invention also relates to a method for preventing and/or treating a disease in a mammal in need thereof or inhibiting SARS-CoV-2 replication or reproduction in a mammal in need thereof, which comprises administering to a mammal in need thereof a prophylactically and/or therapeutically effective amount of the pharmaceutical composition comprising the compound of formula I, its geometric isomer, its pharmaceutically acceptable salt and/or solvate thereof and/or hydrate thereof or the compound of formula I, its geometric isomer, its pharmaceutically acceptable salt and/or solvate thereof and/or hydrate thereof, wherein the disease comprises a SARS-CoV-2 caused disease or infection (e.g., Severe Acute Respiratory Infection (SARI)).

In certain embodiments, the SARS-CoV-2 caused disease or infection described herein includes, but is not limited to, respiratory diseases (e.g., simple infections such as fever, cough, and angina, and the like, pneumonia, acute or severe acute respiratory infections, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, and septic shock, and the like).

The invention also relates to the compound shown in the formula I, a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, which is used for preventing and/or treating diseases or infections caused by SARS-CoV-2 (including but not limited to respiratory diseases (such as simple infections, such as fever, cough, angina and the like, pneumonia, acute or severe acute respiratory tract infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock and the like)).

The invention also relates to the compound shown in the formula I, a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, which is used as a SARS-CoV-2 inhibitor.

The invention also relates to the compound shown in the formula I, a geometric isomer thereof, a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, which is used for inhibiting the replication or propagation of SARS-CoV-2 in cells (such as mammalian cells).

The invention also relates to the pharmaceutical composition, which is used for preventing and/or treating diseases or infections caused by SARS-CoV-2 (including but not limited to respiratory diseases (such as simple infections, such as fever, cough, pharyngalgia and the like, pneumonia, acute or severe acute respiratory infections, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock and the like)).

The invention also relates to said pharmaceutical composition for use as a SARS-CoV-2 inhibitor.

The invention also relates to the pharmaceutical composition for inhibiting the replication or propagation of SARS-CoV-2 in a cell (e.g., a mammalian cell).

In certain embodiments, the SARS-CoV-2 caused disease is COVID-19 according to the invention.

In the present invention, the term "2019 novel coronavirus (2019-nCoV)" is formally classified as Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

In the present invention, the term "2019 novel coronavirus (2019-nCoV) -caused disease" is officially named COVID-19.

In certain embodiments, the mammal comprises a bovine, equine, ovine, porcine, canine, feline, rodent, primate, wherein the preferred mammal is a human, cat, dog, or pig.

The pharmaceutical composition of the present invention can be prepared in various forms according to different administration routes.

According to the present invention, the pharmaceutical composition may be administered in any of the following ways: oral, aerosol inhalation, rectal, nasal, buccal, vaginal, topical, parenteral such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal and intracranial injection or infusion, or via an external reservoir. Among them, oral, intraperitoneal or intravenous administration is preferable.

When administered orally, the compound of formula I, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof may be formulated into any orally acceptable formulation form, including but not limited to tablets, capsules, aqueous solutions or aqueous suspensions. Among them, the carriers generally used for tablets include lactose and corn starch, and additionally, a lubricant such as magnesium stearate may be added. Typical diluents used in capsule formulations include lactose and dried corn starch. Aqueous suspension formulations are generally prepared by mixing the active ingredient with suitable emulsifying and suspending agents. If desired, sweetening, flavoring or coloring agents may be added to the above oral dosage forms.

When administered rectally, the compounds of formula I, their geometric isomers or pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, are generally prepared in the form of suppositories by mixing the drug with a suitable non-irritating excipient. The excipient is in a solid state at room temperature and melts to release the drug at rectal temperature. Such excipients include cocoa butter, beeswax and polyethylene glycols.

When the compound shown in the formula I, the geometrical isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof can be prepared into different forms of topical preparations according to different affected surfaces or organs when the compound is applied topically, particularly for treating affected surfaces or organs which are easy to reach by topical application, such as eyes, skin or lower intestinal tract neurogenic diseases, and the specific description is as follows:

when topically administered to the eye, the compound of formula I, its geometric isomer, or a pharmaceutically acceptable salt thereof, and/or a solvate and/or hydrate thereof, can be formulated in the form of a micronized suspension or solution using sterile saline at a pH that is isotonic, with or without the addition of preservatives such as benzyl chloride alkoxides. In addition, for ophthalmic use, the compounds may also be formulated in the form of ointments such as vaseline.

When applied topically to the skin, the compounds of formula I, their geometric isomers or their pharmaceutically acceptable salts and/or solvates and/or hydrates thereof, can be formulated into suitable ointment, lotion or cream formulations in which the active ingredient is suspended or dissolved in one or more carriers. The carrier that may be used in the ointment herein includes, but is not limited to: mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyethylene oxide, polypropylene oxide, emulsifying wax and water; carriers that can be used in lotions or creams include, but are not limited to: mineral oil, sorbitan monostearate, tween 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

When topically administered to the lower intestinal tract, the compound of formula I, its geometric isomer, or a pharmaceutically acceptable salt thereof, and/or a solvate thereof and/or a hydrate thereof may be formulated in the form of a rectal suppository formulation or a suitable enema formulation as described above, and in addition, a topical transdermal patch may be used.

The compound shown in the formula I, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof can also be used for administration in the form of sterile injection preparations, including sterile injection water or oil suspensions, or sterile injection solutions. Among the carriers and solvents that may be employed are water, ringer's solution and isotonic sodium chloride solution. In addition, the sterilized fixed oil may also be employed as a solvent or suspending medium, such as a monoglyceride or diglyceride.

The medicaments in various dosage forms can be prepared according to the conventional method in the pharmaceutical field.

In the present invention, the term "therapeutically effective amount" or "prophylactically effective amount" refers to an amount sufficient to treat or prevent the disease in a patient, but low enough to avoid serious side effects (at a reasonable benefit/risk ratio), within the scope of sound medical judgment. A prophylactically or therapeutically effective amount of a compound will vary depending on the particular compound selected (e.g., taking into account the potency, effectiveness, and half-life of the compound), the route of administration selected, the disease being prevented or treated, the severity of the disease being prevented or treated, the age, size, weight, and physical condition of the patient being prevented or treated, the medical history of the patient being prevented or treated, the duration of the prevention or treatment, the nature of concurrent therapy, the desired prophylactic or therapeutic effect, and like factors, but can nevertheless be routinely determined by one of ordinary skill in the art.

In addition, the specific dosage and application method of the compound of formula I, its geometric isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate for different patients depends on many factors, including the age, body weight, sex, natural health status, nutritional status, activity intensity of the drug, administration time, metabolic rate, severity of the disease and the subjective judgment of the treating physician. The dosage is preferably between 0.0001-1000mg/kg body weight/day.

Drawings

Figure 1 nafamostat is effective in reducing the viral nucleic acid load on Vero E6 cells infected with SARS-CoV-2 virus, wherein: (a) shows that the nafamostat can inhibit the viral RNA load on cells after the cells are infected with SARS-CoV-2 virus for 48h, the ordinate is the copy number of the viral RNA in the sample, and the abscissa is the drug concentration; (b) nafamostat is shown in the test concentration for 48h with no observed cytotoxicity on test cells, plotted on the ordinate as percent cell viability relative to the negative control (cells only, no drug added) and plotted on the abscissa as drug concentration.

Detailed Description

The following examples are illustrative of preferred embodiments of the present invention and are not to be construed as limiting the invention in any way.