阅读说明：本技术 利托那韦在治疗SARS-CoV-2感染方面的应用 (Application of ritonavir in treating SARS-CoV-2 infection ) 是由 钟武 胡志红 肖庚富 曹瑞源 王曼丽 张磊砢 李薇 樊士勇 李松 于 2020-07-15 设计创作，主要内容包括：本发明涉及利托那韦、其几何异构体及其药学上可接受的盐和/或两者溶剂化物和/或两者水合物,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。(The invention relates to ritonavir, geometric isomers thereof, pharmaceutically acceptable salts thereof and/or solvates thereof and/or hydrates thereof, and a pharmaceutical composition containing the compound, and application of the ritonavir, the geometric isomers thereof, the pharmaceutically acceptable salts thereof and/or the solvates thereof and/or the hydrates thereof in treating SARS-CoV-2 infection.)

1. The use of ritonavir, its geometric isomer or its pharmaceutically acceptable salt and/or solvate and/or hydrate thereof represented by formula I in the preparation of a medicament for preventing and/or treating SARS-CoV-2-caused diseases or infections (e.g. respiratory diseases including but not limited to 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. the ritonavir as 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 is used for preparing the medicine used as the SARS-CoV-2 inhibitor,

3. use of ritonavir, its geometric isomer or its pharmaceutically acceptable salt and/or its solvate and/or its hydrate represented by formula I in the preparation of a medicament for inhibiting the replication or reproduction of SARS-CoV-2 in cells (e.g., mammalian cells),

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

wherein the pharmaceutical composition comprises ritonavir represented by 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. 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 ritonavir represented by 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.

6. 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 (including but not limited to simple infections such as fever, cough, and angina, and the like, pneumonia, acute respiratory infection or severe acute respiratory infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock, and the like), wherein the pharmaceutical composition comprises ritonavir, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof represented by formula I,

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 6, wherein the SARS-CoV-2 caused disease is COVID-19.

8. The use of claim 3 or 5, 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 ritonavir, a geometrical isomer, a pharmaceutically acceptable salt thereof and/or a solvate of the ritonavir and/or a hydrate of the ritonavir, which are shown as a formula I, and a pharmaceutical composition containing the ritonavir and the pharmaceutically acceptable salt, and application of the ritonavir and the pharmaceutical composition to the aspect of treating SARS-CoV-2 infection.

Background

Ritonavir (compound of formula I), the name of british Ritonavir, chemical name 5S- (5R,8R,10R,11R) ] -10-hydroxy-2-methyl-5 (1-methylethyl) -1- [2- (1-methylethyl) -4-thiazolyl ] -3, 6-dioxo-8, 11-bis (phenylmethyl) -2,4,7, 12-tetraazatridecane-13-oic acid, 5-thiazolylmethyl ester is an HIV protease inhibitor, approved for marketing on 29 months of 1999, for use in the treatment of HIV infection.

Ritonavir is a peptide inhibitor of the HIV-1 and HIV-2 proteases. Its inhibition of the HIV protease renders the enzyme unable to regulate the precursor polyprotein responsible for opening-closing, thus generating an infectious inactive immature HIV particle. Ritonavir was studied for antiviral activity in vitro by acute infected lymphoblast cell lines and peripheral blood lymphocytes. Isolated HIV-1 inhibits 50% of viral replication (EC) unlike the cells used₅₀) The concentration of (A) is in the range of 3.8 to 153 nmol.L^-1Mean EC for Low channel clinical isolation of viruses₅₀A value of 22 nmol.L^-1(n＝13)。

Studies reported that during SARS in 2003, poor clinical outcome (ARDS or death) was significantly lower in the treated group following patient use of lopinavir 400mg and ritonavir 100mg than in the control group (2.4% vs 28.8%, p ═ 0.001). However, the researchers believe that lopinavir is a substance inhibiting SARS-CoV, ritonavir does not have the activity of inhibiting SARS-CoV, and the action of ritonavir is to inhibit CYP 3A-mediated lopinavir metabolism, thereby enhancing the serum level of lopinavir (Thorax 2004; 59: 252-. Further literature reports suggest that ritonavir is not effective against SARS-CoV (Biochemical and biological Research Communications 318(2004) 719-725).

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 specific antiviral medicine is available.

Disclosure of Invention

The invention aims to find a medicine with antiviral activity to SARS-CoV-2, which can be used for treating related diseases caused by SARS-CoV-2 infection, such as simple infection, such as fever, cough, pharyngalgia, and the like, pneumonia, acute or severe acute respiratory infection, hypoxic respiratory failure, acute respiratory distress syndrome, sepsis, septic shock, and the like.

The invention discovers that the ritonavir compound shown in the formula I has the function of inhibiting the replication of SARS-CoV-2 and has good potential treatment effect on the aspect of treating diseases caused by SARS-CoV-2 in an in-vitro experiment of SARS-CoV-2 infected cells through creative research.

The present invention relates to compounds having the structure of formula I, geometric isomers thereof, pharmaceutically acceptable salts thereof, and/or solvates thereof, or hydrates thereof:

according to the invention, the pharmaceutically acceptable salts of the compounds shown in the formula I comprise inorganic or organic acid salts and inorganic or organic base salts thereof, and the invention relates to all forms of the 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 the 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 the creative invention and research, the inventor of the invention finds some new action characteristics of the compound shown in the formula I:

the ritonavir compound shown in the formula I can reduce the virus nucleic acid load level of SARS-CoV-2 infected cells under micromolar concentration.

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

the invention also relates to the use of ritonavir shown in formula I, 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 serving as a SARS-CoV-2 inhibitor or in the preparation of a medicament for inhibiting the replication or propagation of SARS-CoV-2 in cells (such as mammalian cells),

the invention also relates to the use of the pharmaceutical composition in the manufacture of a medicament for use as an inhibitor of SARS-CoV-2, or for use in the manufacture of a medicament for use in inhibiting the replication or propagation of SARS-CoV-2 in a cell, such as a mammalian cell,

wherein the pharmaceutical composition comprises ritonavir represented by 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.

The present invention also relates to the 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 (including but not limited to simple infections such as fever, cough, and angina, and the like, pneumonia, acute respiratory infection or severe acute respiratory infection, hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, septic shock, and the like), wherein the pharmaceutical composition comprises ritonavir, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof represented by formula I,

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 present invention also relates to a method for preventing and/or treating a disease or a viral infection in a mammal in need thereof or a method for inhibiting SARS-CoV-2 replication or reproduction in a mammal in need thereof, which comprises administering to the mammal in need thereof a prophylactically and/or therapeutically effective amount of ritonavir represented by formula I, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof or a prophylactically and/or therapeutically effective amount of a pharmaceutical composition comprising a compound of formula I, its geometric isomer or a pharmaceutically acceptable salt thereof and/or a solvate thereof and/or a hydrate thereof,

wherein the disease includes SARS-CoV-2 caused diseases (e.g., respiratory diseases (including but not limited to simple infections such as fever, cough, and angina, pneumonia, acute respiratory infection or Severe Acute Respiratory Infection (SARI), hypoxic respiratory failure and acute respiratory distress syndrome, sepsis, and septic shock, etc.)), and the viral infection includes infection caused by including SARS-CoV-2.

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

preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant.

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

preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or adjuvant.

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

FIG. 1 shows that ritonavir is effective in reducing the viral nucleic acid load on SARS-CoV-2 infected Vero E6 cells. Ritonavir is able to inhibit viral RNA load on cells 48h after infection of cells with SARS-CoV-2 virus, and the inhibitory activity is dose-dependent. In FIG. 1, the left ordinate is the copy number of viral RNA in a sample, the right ordinate is the toxicity of a drug, and the abscissa is the 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.