阅读说明：本技术 硝唑尼特及其活性形式替唑尼特在治疗SARS-CoV-2感染中的应用 (Application of nitazoxanide and its active form tizoxanide in treating SARS-CoV-2 infection ) 是由 钟武 肖庚富 胡志红 王曼丽 曹瑞源 张磊砢 李薇 李月香 赵磊 李松 于 2020-07-15 设计创作，主要内容包括：本发明涉及式I所示硝唑尼特及其活性形式式II所示替唑尼特类化合物,其几何异构体及其药学上可接受的盐和/或其溶剂化物和/或其水合物,及含有此化合物的药物组合物,用于预防和/或治疗冠状病毒(例如SARS-CoV-2)的感染,<Image he="199" wi="700" file="DDA0002586001900000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The invention relates to nitazoxanide shown in formula I and tizoxanide compound shown in formula II in active form, geometric isomer, pharmaceutically acceptable salt and/or solvate and/or hydrate thereof, and pharmaceutical composition containing the compound, which is used for preventing and/or treating infection of coronavirus (such as SARS-CoV-2),)

1. The use of a compound of formula I and/or formula II, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, for the manufacture of a medicament for the prevention and/or treatment of a disease or infection caused by a coronavirus (e.g. SARS-CoV-2), including but not limited to respiratory diseases (e.g. simple infections such as fever, cough and angina etc., 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 a coronavirus, such as SARS-CoV-2, for example a respiratory disease, such as a simple infection like fever, cough, and sore throat etc., pneumonia, acute or severe acute respiratory infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis and septic shock etc, wherein the pharmaceutical composition comprises a compound of formula I and/or formula II, 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.

3. A compound shown as a formula I and/or a formula II,

The use of a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof for the manufacture of a medicament for use as an inhibitor of coronavirus (e.g. SARS-CoV-2).

4. A compound shown as a formula I and/or a formula II,

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

5. Use of a pharmaceutical composition comprising a compound of formula I and/or formula II, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof for the manufacture of a medicament for use as an inhibitor of a coronavirus, such as SARS-CoV-2,

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.

6. Use of a pharmaceutical composition for the manufacture of a medicament for inhibiting replication or propagation of a coronavirus (e.g., SARS-CoV-2) in a cell (e.g., a mammalian cell), wherein the pharmaceutical composition comprises a compound of formula I and/or formula II, 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 4 or 6, wherein the mammal comprises a bovine, equine, ovine, porcine, canine, feline, rodent, primate, such as a human, feline or porcine.

Technical Field

The invention relates to a compound nitazoxanide shown in the following formula I and an active form of the compound nitazoxanide shown in the formula II, geometrical isomers of the nitazoxanide and the active form of the nitazoxanide, pharmaceutically acceptable salts of the nitazoxanide and the active form of the nitazoxanide, and/or solvates and/or hydrates of the nitazoxanide and a pharmaceutical composition containing the compound, and application of the nitazoxanide and the salt in the aspect of treating SARS-CoV-2 infection.

Background

Nitazoxanide (compound of formula I), chemical name is ortho [ N- (5-nitrothiazol-2-yl) carbamoyl ] phenol acetate (2- (acetoxy) -N- (5-nitro-2-thiazole) is a thiazole drug, which was first synthesized in the seventies of the last century (Rossinol, J.F., Cavier, R.,1975.Synthesis and antiparastic activity of 2-benzamidonitione, chem.Abstract.83, 28216n.). Nitazoxanide is a prodrug that hydrolyzes in vivo shortly after administration to its active metabolite, Tizoxanide (TIZ, compound of formula II), and thus exerts its pharmacological effects.

Nitazoxanide is initially researched as an oral antiparasitic drug, has good antiprotozoal activity in vitro and different animal models, and is clinically researched for treating human intestinal tapeworm for the first time. Subsequent in vitro and in vivo researches find that the compound has good inhibition effect on new protozoa and cryptosporidium. In 1998, the FDA in the united states approved nitazoxanide as a clinical treatment for infections with cryptosporidium parvum and giardia in adults and children over one year of age. In 2006, clinical studies confirmed that nitazoxanide can effectively treat diarrhea in children caused by rotavirus infection. In addition, Nitazoxanide has also been shown to have a good clinical therapeutic effect on adult viral gastroenteritis and hepatitis C (Rosssignol JF, Nitazoxanide: afirst-in-class branched-specific Antiviral agent, Antiviral Res.2014; 110: 94-103.).

In recent years, multiple laboratory researches prove that nitazoxanide has broad-spectrum antiviral activity, and a series of laboratory researches show that nitazoxanide can block the maturation of influenza virus hemagglutinin protein at the post-translational stage and has high-efficiency inhibitory activity on influenza A and influenza B. Clinical study results show that nifedipine can effectively reduce the course of common influenza by taking 600mg twice a day for 5 days (Haffizulla J, Hartman A, Hoppers M, et al, effective of nitrozonides in adults and adolescents with access to uncompounded inflenza: available-blind, randomised, placode-controlled, phase 2b/3trial, Lancet InfectDis.2014; 14(7): 609-18.).

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 virus infection are mainly pneumonia, and can be divided 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 targeted antiviral medicine is available.

Disclosure of Invention

The present invention aims at finding medicine with antiviral activity to SARS-CoV-2 and may be used in the treatment of SARS-CoV-2 infection. The invention discovers that nitazoxanide shown in formula I has the functions of protecting SARS-CoV-2 infected cells and inhibiting SARS-CoV-2 replication, and has good potential treatment effect in the aspect of treating SARS-CoV-2-caused diseases through creative research.

The invention relates to thiazole compounds shown in a formula I or a formula II, geometrical isomers thereof, or pharmaceutically acceptable salts thereof and/or solvates thereof or hydrates thereof:

in certain embodiments, the pharmaceutically acceptable salts of the compounds of the present invention include inorganic or organic acid salts thereof, as well as inorganic or organic base salts, 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.

According to the invention, the compound shown in formula I or formula II can protect cytopathic effect (CPE) of cells caused by SARS-CoV-2 virus infection, inhibit SARS-CoV-2 virus replication on cells and reduce SARS-CoV-2 virus nucleic acid load in cell culture.

After creative invention and research, the inventor of the invention finds out a new action characteristic of the compound shown as the formula I or the formula II in cells: the compound of formula I or formula II can reduce the viral nucleic acid load level of SARS-CoV-2 virus infected cells at micromolar concentrations.

The invention also relates to a compound shown in the formula I and/or the formula II, a geometric isomer thereof or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof, in the preparation of a medicament for preventing and/or treating diseases or infections (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 tract infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock and the like)) caused by the coronavirus (such as SARS-CoV-2),

the invention also relates to application of the compound shown in the formula I and/or the formula II, geometric isomers thereof or pharmaceutically acceptable salts thereof and/or solvates thereof and/or hydrates thereof in preparing medicaments serving as coronavirus (such as SARS-CoV-2) inhibitors.

The invention also relates to application of the compound shown in the formula I and/or the formula II, geometric isomer thereof or pharmaceutically acceptable salt thereof and/or solvate thereof and/or hydrate thereof in preparing a medicament for inhibiting replication or propagation of coronavirus (such as 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 and/or the formula II, the geometrical isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the 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 the application of the pharmaceutical composition containing the compound shown in the formula I and/or the formula II, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof or the compound shown in the formula I and/or the formula II, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing medicines for treating respiratory system diseases (simple infection 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 a method of preventing and/or treating a disease or inhibiting the replication or propagation of a coronavirus (e.g., SARS-CoV-2) in a mammal in need thereof, the method 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 and/or formula II, a geometric isomer thereof, or a pharmaceutically acceptable salt thereof, and/or a solvate thereof, and/or a hydrate thereof, or the compound of formula I and/or formula II, a geometric isomer thereof, or a pharmaceutically acceptable salt thereof, and/or a solvate thereof, and/or a hydrate thereof, wherein the disease comprises a disease or infection (e.g., Severe Acute Respiratory Infection (SARI)) caused by a coronavirus (e.g., SARS-CoV-2).

In certain embodiments, the disease is a SARS-CoV-2 caused disease or infection.

In certain embodiments, the SARS-CoV-2 caused disease includes, but is 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, septic shock, etc.).

The invention also relates to application of the pharmaceutical composition containing the compound shown in the formula I and/or the formula II, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing a medicament for preventing and/or treating diseases or infections (such as 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)) caused by the coronavirus (such as SARS-CoV-2).

The invention also relates to application of the pharmaceutical composition containing the compound shown in the formula I and/or the formula II, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing a medicament serving as a coronavirus (such as SARS-CoV-2) inhibitor.

The invention also relates to application of the pharmaceutical composition containing the compound shown in the formula I and/or the formula II, the geometric isomer thereof or the pharmaceutically acceptable salt thereof and/or the solvate thereof and/or the hydrate thereof in preparing a medicament for inhibiting replication or propagation of coronavirus (such as SARS-CoV-2) in cells (such as mammalian cells).

The invention also relates to the compound shown in the formula I and/or the formula II, a geometric isomer thereof or 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 (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 tract infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock and the like)) caused by the coronavirus (such as SARS-CoV-2).

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

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

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

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

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 the present invention, the pharmaceutical composition may 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 therapeutically or prophylactically effective amount of a compound will vary depending upon factors such as the particular compound selected (e.g., in view of the potency, effectiveness, and half-life of the compound), the route of administration selected, the disease being treated or prevented, the severity of the disease being treated or prevented, the age, size, weight, and physical condition of the patient being treated or prevented, the medical history of the patient being treated or prevented, the duration of treatment or prevention, the nature of concurrent therapy, the desired therapeutic or prophylactic effect, and the like, 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 shows that nitazoxanide is effective in reducing viral nucleic acid load on Vero E6 cells infected with SARS-CoV-2, where (a) nitazoxanide is able to suppress viral RNA load on cells after SARS-CoV-248 h infection of cells, with ordinate being the copy number of viral RNA in the sample and abscissa being the drug concentration; (b) it is shown that no cytotoxicity was observed with nitazoxanide at the tested concentration for 48h, with percent cell viability relative to the negative control (cells only, no drug) on the ordinate and drug concentration on the abscissa.

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.