WO2021047437A1

WO2021047437A1 - Pharmaceutical composition for treating viral influenza and preparation thereof

Info

Publication number: WO2021047437A1
Application number: PCT/CN2020/113105
Authority: WO
Inventors: 陈小新; 李海军; 刘卓伟; 刘志强; 万春喜; 龙超峰
Original assignee: 广东众生睿创生物科技有限公司
Priority date: 2019-09-10
Filing date: 2020-09-02
Publication date: 2021-03-18
Also published as: CN114502150B; CN114502150A

Abstract

Disclosed are a pharmaceutical composition and a preparation thereof, wherein the pharmaceutical composition contains two effective ingredients with a mass ratio being in a certain range. The pharmaceutical composition has a significantly better anti-influenza virus effect, and after a comprehensive evaluation, the pharmaceutical composition is considered to have good prospects for preparing drugs.

Description

Medicinal composition and preparation for treating viral cold

Technical field

The invention belongs to the field of pharmaceutical preparations, and particularly relates to a pharmaceutical composition for treating viral influenza and a preparation containing the pharmaceutical composition.

Background technique

Influenza virus, known as influenza virus (IFV), is a segmented single-stranded antisense RNA virus that can cause influenza in humans and animals. According to the statistics of the World Health Organization (WHO), the annual seasonal epidemic of influenza can cause 3 million to 5 million severe cases and 290,000 to 650,000 deaths worldwide. Every influenza pandemic has a huge impact on global public health and the economy. Devastation.

The current clinical plan for influenza includes vaccination and antiviral drugs for chemotherapy and chemoprevention. One of the important antiviral drugs is neuraminidase inhibitors, such as Oseltamivir (OSE), Zana Zanamivir, Peramivir, etc., such antiviral drugs have obvious effects on influenza A virus, and are commonly used clinical drugs for the treatment of viral influenza. However, similar to antibiotic products, with the long-term use of antiviral products, the resulting drug resistance has become increasingly serious. According to the monitoring data of the WHO Western Pacific region, the proportion of oseltamivir-resistant strains of the seasonal A H1N1 influenza virus in China from 2008 to 2009 has reached 28%, and the resistance of influenza viruses affects existing anti-influenza viruses. The therapeutic effect of the drug.

Among them, after oral administration, oseltamivir is metabolized into its active metabolite oseltamivir carboxylic acid to exert its drug effect. By competitively binding with the active site of influenza virus neuraminidase, it interferes with the virus from the infected host cell. During the release process, thereby reducing the spread of influenza A or B viruses.

Patent WO2018041263 discloses a series of pyrimidine derivatives. In vitro activity data showed that some compounds showed positive effects in the test of inhibiting influenza virus replication. In further animal experiments, some compounds also showed significant therapeutic effects on influenza A virus H1N1 mouse infection model. Among them, compound 1 The comprehensive performance of (Example 4, WX-216) is relatively outstanding, and it is considered to have a better prospect of preparing medicines.

On the other hand, influenza virus has the characteristics of high mutation rate and multiple inter-virus recombination phenomena. Although new structures and new mechanisms of active compounds are constantly being discovered, in the long run, the use of a single antiviral drug may not be effective in a short time. Inhibiting/reducing the concentration of the virus in the body and curing the defects of the viral cold, the corresponding increase in the dosage and the lengthening of the treatment time also bring a greater risk of medication to the patient. Finberg RW et al., J Infect Dis, 2019 Mar 15; 219(7): 1026-1034. A combination experiment of Pimodivir and oseltamivir was published, and the results showed that the combination of 600 mg Pimodivir and 75 mg oseltamivir Use can minimize the load of influenza A virus.

Therefore, finding new solutions to solve the drug resistance problem of new clinically promising products, reducing the probability of adverse drug reactions, and responding to the huge demand for anti-influenza virus drugs that are currently unmet clinically is an urgent need for existing technologies to solve technical problem.

Summary of the invention

The first object of the present invention is to overcome at least one of the above-mentioned shortcomings of the prior art and provide a pharmaceutical composition for the treatment of viral influenza, which utilizes the synergistic effect between the active ingredients to solve the problem of influenza Technical problems such as the drug resistance of the virus can effectively reduce the risk of clinical medication. After comprehensive evaluation, it is believed that the pharmaceutical composition has a better prospect for preparing a medicine.

The purpose of the present invention is achieved through the following technical solutions:

A pharmaceutical composition comprising a first active ingredient and a second active ingredient, wherein the first active ingredient is compound 1, its corresponding ester, its corresponding salt, its corresponding ester salt, or a combination thereof The second effective ingredient is a neuraminidase inhibitor; the mass ratio of the first effective ingredient to the second effective ingredient is 1-8:1.

The first active ingredient refers to a mixture obtained by mixing one or more of the compound 1, the ester corresponding to the compound 1, the salt of the compound 1, and the salt of the ester corresponding to the compound 1, in any ratio.

The compound 1, the ester corresponding to the compound 1, the salt of the compound 1, and the salt of the ester corresponding to the compound 1 each include the subordinate anhydrate, ansolvate, hydrate, and solvate thereof.

The ester corresponding to compound 1 refers to the ester formed by compound 1 and organic acid, including but not limited to methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl and the like of compound 1.

The salt of compound 1 is the salt formed by compound 1 and organic acid and/or inorganic acid, or the salt formed by compound 1 and organic base and/or inorganic base, including but not limited to the hydrochloride and hydrobromic acid of compound 1. Salt, sulfate, phosphate, benzenesulfonate, p-toluenesulfonate, methanesulfonate, tartrate, camphorsulfonate, lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, aluminum salt, Ammonium salt, ethylenediamine salt, triethylamine salt, etc.

The salt of the corresponding ester of compound 1 refers to the salt of the ester formed by the aforementioned compound 1 with an organic acid and an organic base and/or an inorganic base and/or an organic acid and/or an inorganic acid, including but not limited to the salt of the methyl ester of compound 1 Acid salt, compound 1 methyl ester sulfate, compound 1 ethyl ester hydrochloride, compound 1 ethyl ester sulfate, etc.

The neuraminidase inhibitor referred to by the second active ingredient is oseltamivir (Oseltamivir) or its derivatives, zanamivir (zanamivir) or its derivatives, peramivir (Peramivir) or its derivatives A mixture obtained by mixing one or two or more of the substances in any ratio; the derivatives of Oseltamivir, zanamivir, and Peramivir are independently selected from them Acids and/or bases can also be esters, amides, or other derivatives such as carboxylic acids and their salts obtained after hydrolysis, and each includes its subordinate anhydrates, ansolvates, hydrates and solvents For example, oseltamivir or its derivatives can be selected as oseltamivir free state, or oseltamivir phosphate, oseltamivir carboxylic acid, oseltamivir carboxylate, and the like.

The mass ratio of the first effective ingredient to the second effective ingredient is 1 to 8:1, preferably, the mass ratio of the first effective ingredient to the second effective ingredient is 100 to 600:75; specifically, the The mass ratio of the first active ingredient to the second active ingredient is about 100:75, 200:75, 300:75, 400:75, 500:75, 600:75.

Unless otherwise specified, the quality of the salt in the present invention refers to the mass based on free base/acid equivalent, and the hydrate/solvate refers to the mass based on dry matter (on an equivalent basis). Anhydrous basis), the mass of metabolites (such as ester hydrolysis to obtain carboxylic acid) refers to the mass of the compound before metabolism.

In a preferred technical scheme of the present invention, the first active ingredient in the pharmaceutical composition is compound 1, and the second active ingredient is oseltamivir phosphate; the combination of compound 1 and oseltamivir phosphate in the pharmaceutical composition The mass ratio is 100-600:75; specifically, the mass ratio of the compound 1 to oseltamivir phosphate is about 100:75, 200:75, 300:75, 400:75, 500:75, 600:75.

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is the sodium salt of compound 1, and the second active ingredient is oseltamivir phosphate; the sodium salt of compound 1 in the pharmaceutical composition is combined with oseltamivir phosphate. The mass ratio of setavir phosphate is 100-600:75; specifically, the mass ratio of the sodium salt of compound 1 to oseltamivir phosphate is about 100:75, 200:75, 300:75, 400:75, 500:75, 600:75 .

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is compound 1 hydrochloride, and the second active ingredient is oseltamivir phosphate; in the pharmaceutical composition, compound 1 hydrochloride The mass ratio with oseltamivir phosphate is 100-600:75; specifically, the mass ratio of the hydrochloride of compound 1 to oseltamivir phosphate is about 100:75, 200:75, 300:75, 400:75, 500: 75,600:75.

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is the potassium salt of compound 1, and the second active ingredient is oseltamivir phosphate; The mass ratio of setavir phosphate is 100-600:75; specifically, the mass ratio of the potassium salt of compound 1 to oseltamivir phosphate is about 100:75, 200:75, 300:75, 400:75, 500:75, 600:75 .

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is the calcium salt of compound 1, and the second active ingredient is oseltamivir phosphate; The mass ratio of setavir phosphate is 100-600:75; specifically, the mass ratio of the calcium salt of compound 1 to oseltamivir phosphate is about 100:75, 200:75, 300:75, 400:75, 500:75, 600:75 .

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is p-toluenesulfonate of compound 1, and the second active ingredient is oseltamivir phosphate; The mass ratio of tosylate to oseltamivir phosphate is 100-600:75; specifically, the mass ratio of p-toluenesulfonate to oseltamivir phosphate of the compound 1 is about 100:75,200: 75,300:75,400:75,500:75,600:75.

The inventor surprisingly found in experiments that the foregoing contains the first active ingredient (compound 1, its corresponding ester, its corresponding salt, its corresponding ester salt or a combination thereof) and the second active ingredient (neuraminidase When the mass ratio of the two is in a specific range, the drug composition of inhibitor) has a better drug synergistic effect, showing a significantly better anti-influenza virus effect than a single component, and also showing a better anti-influenza virus effect than the aforementioned specific range. The pharmaceutical composition with the external mass ratio has significantly better anti-influenza virus effect. It can be known that when the mass ratio of the first active ingredient and the second active ingredient in the unit preparation is within the specific range, it has better anti-influenza virus effect. Effect: After comprehensive evaluation of other pharmaceutical properties (such as mixture stability, fluidity, etc.), it is believed that the pharmaceutical composition has a better prospect for pharmaceutical preparation.

Another object of the present invention is to disclose a preparation method of the aforementioned pharmaceutical composition, which can ensure the stable preparation of the aforementioned pharmaceutical composition. Specifically, the preparation method adopts a conventional mixing method in the art; more specifically, the preparation method may be a direct mixing method, an equal incremental method, etc., and the mixing equipment used may be a common mixing method in the field, depending on the preparation scale. Equipment, such as V-type mixer, double cone mixer, rotary mixer, etc., can also be prepared by manual mixing in small-scale preparation.

The third object of the present invention is to disclose a preparation containing the aforementioned pharmaceutical composition. Specifically, the preparation includes the aforementioned pharmaceutical composition and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is selected from any one or two or more of fillers, binders, disintegrants, glidants, lubricants, and flavoring agents.

Specifically, the filler is selected from calcium carbonate, magnesium carbonate, calcium phosphate, calcium sulfate, magnesium oxide, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, sucrose, lactose, fructose, xylitol, mannitol , Starch or its derivatives, dextrin, microcrystalline cellulose, etc., a mixture obtained by mixing any one or two or more of them in any ratio.

Specifically, the binder is selected from gum arabic, gelatin, tragacanth, dextrin, polyvinylpyrrolidone, starch and its derivatives, sodium alginate, sorbitol, syrup, hydroxypropyl methylcellulose, methyl cellulose Any one or two of cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose and polymethacrylate, etc. The mixture obtained by mixing more than one species in any ratio.

Specifically, the disintegrant is selected from starch, alginic acid, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, croscarmellose sodium, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropyl methyl A mixture obtained by mixing any one or two or more of base cellulose, microcrystalline cellulose, and methyl cellulose in an arbitrary ratio.

Specifically, the glidant is a mixture obtained by mixing any one or more of colloidal silicon dioxide, powdered cellulose, magnesium trisilicate, silicon dioxide, and talc in any ratio.

Specifically, the lubricant is selected from calcium stearate, glyceryl monostearate, glyceryl palmitostearate, magnesium stearate, microcrystalline cellulose, sodium benzoate, sodium chloride, and lauryl sulfate A mixture obtained by mixing any one or two or more of sodium, sodium stearyl fumarate, talc, zinc stearate, and polyethylene glycol in an arbitrary ratio.

Specifically, the flavoring agent is selected from any one or a mixture of two or more of stevioside, aspartame, and other flavors and sweeteners commonly used in the art in any ratio.

The preparation containing the aforementioned pharmaceutical composition is an oral preparation, specifically a powder, granule, pellet, capsule, tablet, solution or lozenge.

Based on the beneficial effects of the aforementioned pharmaceutical composition, such as drug synergy, etc., the preparations containing the aforementioned pharmaceutical composition also exhibit significantly better anti-influenza virus effects than those containing single-component preparations, and also show greater anti-influenza virus effects than those containing the aforementioned specific range. The pharmaceutical composition preparations with other mass ratios have significantly better anti-influenza virus effects, and have a better market prospect.

Compared with the prior art, the present invention has the following outstanding advantages and beneficial effects:

1. A pharmaceutical composition is provided, which comprises a first active ingredient (compound 1, its corresponding ester, its corresponding salt, its corresponding ester salt or a combination thereof) with a mass ratio in a specific range, and The second active ingredient (neuraminidase inhibitor), which has a better drug synergistic effect, in which the neuraminidase inhibitor is hydrolyzed with oseltamivir and/or oseltamivir salt and/or oseltamivir The oseltamivir carboxylic acid or oseltamivir carboxylate obtained later has the best effect, and after comprehensive evaluation, it is believed that the pharmaceutical composition has a better prospect for preparing medicines.

2. A preparation method of the pharmaceutical composition is provided, which can ensure the stable preparation of the aforementioned pharmaceutical composition.

3. A preparation containing the aforementioned pharmaceutical composition is provided, which correspondingly shows a significantly better anti-influenza virus effect than a preparation containing a single component, and is also shown to be more effective than a pharmaceutical combination containing a mass ratio outside the aforementioned specific range The drug preparation has significantly better anti-influenza virus effect and has a better market prospect.

The contribution of the present invention lies in the discovery that a combination of drugs within a specific range has a synergistic effect. Therefore, those skilled in the art can understand that when the combination of drugs in a unit dosing unit is within the scope of the present invention, it can be considered to be used. The technical solution protected by the present invention; specifically, the aforementioned unit administration unit refers to the smallest unit for clinical use, such as: unit tablet, unit capsule, unit bottle of oral liquid, unit package of granules, etc. .

detailed description

The present invention will be further described in detail below in conjunction with examples, but the implementation of the invention is not limited thereto.

The oseltamivir phosphate used in the examples is the bulk drug of the commercially available Tamiflu, CAS: 204255-11-8, with a purity of >99%; the oseltamivir used is the free substance of the commercially available Tamiflu bulk drug, CAS ：196618-13-0, purity>99%; the oseltamivir carboxylic acid used is a commercially available intermediate, CAS: 187227-45-8, purity>99%.

The zanamivir (CAS: 139110-80-8) and peramivir (CAS: 229614-55-5) used in the examples are the raw materials of the commercially available preparations with a purity of >99%.

Example 1 Preparation of compound 1

The compound 1 was prepared by the method disclosed in Example 4 of WO2018041263, and the structure characterization proved that the obtained product was compound 1.

Example 2 Preparation of the hydrochloride salt of compound 1

Take 5g of compound 1 into a 250mL eggplant-shaped flask, add THF (100mL), add 0.98mL of hydrochloric acid (concentrated hydrochloric acid dissolved in 9mL THF), stir for 12 hours at 30°C, filter the solid, and dry the filter cake at 40°C in vacuo to obtain compound 1. The hydrochloride.

Example 3 Preparation of p-toluenesulfonate of compound 1

Take 5g of compound 1 into a 250mL eggplant-shaped flask, add THF (100mL), add p-toluenesulfonic acid monohydrate (2.26g, dissolved in 10mL THF), stir at 30°C for 12 hours, filter the solids, and vacuum the filter cake at 40°C After drying, a solid (0.425 g) was obtained. This solid (0.101g) was added to acetone (2mL) and slurried for 12h to obtain the p-toluenesulfonate of compound 1.

Example 4 Preparation of the sodium salt of compound 1

Take 5g of compound 1 into a 250mL eggplant-shaped flask, add THF (100mL), add NaOH aqueous solution (0.477g, dissolved in 1mL of water), stir for 12 hours at 30°C, filter the solid, and dry the filter cake at 40°C under vacuum to obtain compound 1 The sodium salt.

Example 5 Preparation of the potassium salt of compound 1

Weigh 2g of compound 1 into a 100mL eggplant-shaped flask, add THF (35mL), add KOH aqueous solution (0.255g, dissolved in 0.5mL water and 5mL THF), stir for 12 hours at 30°C, filter the solids, and vacuum the filter cake at 40°C After drying, the potassium salt of compound 1 was obtained.

Example 6 Preparation of calcium salt of compound 1

Take 2g of compound 1 into a 100mL eggplant-shaped flask, add THF (35mL), add calcium hydroxide aqueous solution (0.168g, dissolved in 0.5mL water and 5mL THF), stir for 12 hours at 25°C, filter the solids, and filter cake at 40°C Dry under vacuum to obtain a solid (1.440g), dissolve this solid (0.204g) in a mixed solvent of ethanol and water (ethanol:water=3:1) (4mL) to make a slurry, stir at 25°C for 12 hours, and filter the solid , The filter cake was vacuum dried at 40°C to obtain the calcium salt of compound 1.

Example 7

Take 200 g of compound 1 prepared in Example 1, and mix it with 75 g of oseltamivir phosphate (calculated as oseltamivir) uniformly to obtain a pharmaceutical composition with a mass ratio of 200:75.

Example 8

Using the same method as in Example 7, the mass ratios of compound 1 and oseltamivir phosphate were prepared respectively as 50:75, 100:75, 300:75, 400:75, 500:75, 600:75, 700: 75 pharmaceutical compositions.

Using the same method as in Example 7, the mass ratios of the salt of compound 1 prepared in Examples 2-6 and oseltamivir or oseltamivir phosphate were 50:75 and 100:75, respectively. , 200:75, 300:75, 400:75, 500:75, 600:75, 700:75 pharmaceutical composition, the mass of the salt refers to the mass of the free substance (free base/acid equivalent) .

Example 9 In vitro activity comparison experiment

1. The purpose of the experiment

Taking compound 1 and oseltamivir carboxylic acid (active metabolite) as the research object, analyze the synergy of compound 1 and oseltamivir (or oseltamivir carboxylic acid) in combination with influenza A virus at a certain ratio Inhibition, combined with the plasma protein binding rate of the two, convert and predict its combined effect in vivo.

2. Experimental materials

Virus: A/Aichi/2/1968 (H3N2), purchased from ATCC.

Cells: Dog kidney epithelial cell line (Madin-Darby Canine Kidney Cells, MDCK).

Drug dissolution method: Compound 1 and the active metabolite of oseltamivir (oseltamivir carboxylic acid, calculated as oseltamivir) were dissolved in dimethyl sulfoxide (DMSO), prepared as a 100mM/L mother liquor, and stored for later use; Dilute with Medium Essential Medium (MEM) as a working solution (Note: the final concentration of DMSO is 1%).

3. Experimental method

MDCK cells were seeded into a 96-well plate at a concentration of about 2.5×10 ⁴ per well. After the cells grew into a monolayer (about 24 hours), the culture medium was discarded, and the cell surface was washed twice with PBS. Add 100μL/well of virus diluent containing MOI=0.01, incubate in a 37°C, 5% CO ₂ incubator for 2 hours, discard the virus solution, add compound 1 and oseltamivir carboxylic acid to two-fold dilution to different concentrations and 0nM Combination medication (ie, compound 1 is: 1, 0.5, 0.25, 0.125, 0.0625, 0.03125, 0.015625, 0 nM; oseltamivir carboxylic acid is: 0.25, 0.125, 0.0625, 0.03125, 0 μM, respectively). Add 100μL/well of the combined drug to 96-well cells, set up 3 replicate wells for each concentration, and set up normal cell group, virus control, and drug control group at the same time; _{incubate in a 37℃, 5% CO 2} incubator, and observe the cells every day The condition of the disease, and then use MTT to detect the cell viability of each well, and the results of virus inhibition after 72 hours of observation are shown in Table 1 below.

Table 1 Compound 1 combined with oseltamivir carboxylic acid anti-A/Aichi/2/1968 (H3N2) (inhibition rate %)

It can be seen from the above data that when the molar ratio of compound 1: oseltamivir carboxylic acid is 0.5nM:0.03125μM to 0.015625

When nM: 0.25μM (1:62.5～16000), it shows a significant synergistic inhibitory effect on the tested influenza virus, and the specific manifestation is that the virus inhibition rate after the combination is higher than the single-agent virus inhibition rate; in view of compound 1 After entering the human body, the plasma protein binding rate can reach 99.9%. It can be seen that the compound 1 in the free state in the body accounts for about 0.1%, and the binding rate of oseltamivir carboxylic acid in the human plasma protein is 0.3%, which shows that the body is in a free state. Oseltamivir carboxylic acid accounts for about 99.7%; after conversion, that is, in the body, the molar ratio of compound 1 to oseltamivir carboxylic acid should be about 1:16 to 16:1 when there is a good synergistic effect. That is, when the mass ratio of compound 1 to oseltamivir is approximately in the range of 1:11.3 to 22.5:1 (440:4992 to 7040:312), there is a good synergistic effect.

In follow-up experiments, it was found that compound 1 has a synergistic effect with oseltamivir, zanamivir and peramivir, but compared with other groups, the oseltamivir group has a synergistic performance significantly better than other groups.

Example 10 In vivo activity experiment

Study on the efficacy of drugs on PR8 influenza virus

1. Research on the protective effect of drugs on the death of PR8 influenza virus (provide PR8 virus 2h in advance)

Cultivate influenza virus with higher titer, and infect BALB/C mice with 2LD50 (lethal dose) intranasally.

Continuous 5-day administration group: 2 hours after infection, intragastric administration, twice a day, continuous intragastric administration for 5 days, set the drug group compound 1: 30 mg/kg, compound 1: 20 mg/kg, compound 1: 5 mg/kg, Compound 1: 1.5 mg/kg, oseltamivir phosphate: 10 mg/kg, oseltamivir phosphate + compound 1 = 10 mg/kg + 30 mg/kg, oseltamivir phosphate + compound 1 = 10 mg/kg +20mg/kg, oseltamivir phosphate + compound 1 = 10 mg/kg + 5 mg/kg, oseltamivir phosphate + compound 1 = 10 mg/kg + 1.5 mg/kg; and set a blank control group and virus control Groups, 10 mice in each group; observe from the first day of virus infection and continuously observe for 14 days. Observe and record the weight change and death of the infected mice every day, and calculate according to the following formula:

Mortality rate (%) = number of dead mice in each group (only)/total number before infection (only) × 100%

The results showed that after the virus infection, the mice developed symptoms such as shortness of breath, curled up and trembling, weight loss, and died; after 14 days of continuous observation, the mice in the blank group were in good condition and no death was observed, while the mortality of the mice in the virus group reached 100%; compound 1:30mg/kg group has a complete protective effect on infected mice, and its mortality rate is 0%; compound 1:20mg/kg group has obvious protective effect on infected mice, and its mortality rate is 20%; compound The mortality of mice in the 1:5 mg/kg administration group was 80%; the mortality of mice in the oseltamivir phosphate: 10 mg/kg administration group and the compound 1:1.5 mg/kg administration group was 100%. After compound 1 was used in combination with oseltamivir phosphate at the doses of 5 mg/kg and 20 mg/kg, the mortality rate was significantly reduced, and the synergistic effect was obvious. The results are shown in Table 2:

Table 2 Mortality and survival rate of mice in each group

It can be seen that the combination of 5 mg/kg, 20 mg/kg compound 1 and 10 mg/kg oseltamivir phosphate showed a good synergistic antiviral effect on the mouse pharmacodynamic model. Based on the conversion of compound 1 and oseltamivir phosphate in different species (mice and healthy people) pharmacokinetic test results, administration of 5mg/kg, 20mg/kg of compound 1 to mice and 75mg, 600mg of human body exposure Similar, and the exposure of 10 mg/kg oseltamivir phosphate to mice is similar to that of 75 mg of human body active metabolite exposure. It can be seen that the combined use of 600, 75 mg of compound 1 and 75 mg of oseltamivir phosphate is effective, indicating that it is a compound 1 When the mass ratio of oseltamivir phosphate is 75-600:75 (1-8:1), the composition has a better synergistic effect in vivo.

Based on the above, it can be seen that the synergistic effect of the composition of the present invention is inconsistent in vivo and in vitro; specifically, the composition exhibits inconsistent in vivo and in vitro regardless of the range of drug synergistic effects and the more effective range. The specific performance is that the mass ratio of oseltamivir and compound 1 is about 1:22.5 to 11.3:1 according to the range of drug synergy in vitro, and the range of drug synergy is surprisingly large in in vivo experiments. The reduction is that the mass ratio of compound 1 and oseltamivir phosphate is 75～600:75 (1～8:1), which is synergistic; further, the mass ratio of compound 1 and oseltamivir phosphate is 75: 75,100:75,200:75,300:75,400:75,500:75,600:75, it shows a good synergy of drugs in the body;

Further expanded experiments have shown that when compound 1, its corresponding ester, its corresponding salt, its corresponding ester salt or a combination thereof, and oseltamivir or oseltamivir phosphate also show basically the same performance, Among them, the pharmaceutical composition of compound 1 and oseltamivir phosphate, and the pharmaceutical composition of compound 1 sodium salt and oseltamivir phosphate have the best effect, and they are more effective within the scope of the patent. The range of viruses all have better drug synergy in vivo, so they are considered to have better clinical prospects.

Example 11

The pharmaceutical composition obtained in Example 7 was mixed with an appropriate amount of filler, binder, and disintegrant, and then directly compressed to prepare a tablet (200mg+75mg).

The above-mentioned embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, etc. made without departing from the spirit and principle of the present invention Simplified, all should be equivalent replacement methods, and they are all included in the protection scope of the present invention.

Claims

A pharmaceutical composition, characterized in that the pharmaceutical composition comprises a first active ingredient and a second active ingredient, wherein the first active ingredient is compound 1, its corresponding ester, its corresponding salt, and its corresponding ester salt Or a combination thereof; the second active ingredient is a neuraminidase inhibitor; the mass ratio of the first active ingredient to the second active ingredient is 1-8:1
The pharmaceutical composition according to claim 1, wherein the first active ingredient is any one or two or more of compound 1, the ester corresponding to compound 1, the salt of compound 1, and the salt of ester corresponding to compound 1. Mix the resulting mixture in proportions.
The pharmaceutical composition according to claim 2, wherein the ester corresponding to compound 1 refers to the ester formed by compound 1 and organic acid, preferably the methyl, ethyl, n-propyl, isopropyl, n-propyl of compound 1 Butyl ester, tert-butyl ester; the salt of compound 1 is the salt formed by compound 1 and organic acid and/or inorganic acid, or the salt formed by compound 1 and organic base and/or inorganic base, preferably hydrochloride, hydrobromide Acid salt, sulfate, phosphate, benzenesulfonate, p-toluenesulfonate, methanesulfonate, tartrate, camphorsulfonate, lithium salt, sodium salt, potassium salt, calcium salt, magnesium salt, aluminum salt , Ammonium salt, ethylenediamine salt, triethylamine salt; the corresponding ester salt of compound 1 refers to the ester formed by compound 1 and organic acid with organic base and/or inorganic base and/or organic acid and/or inorganic acid The formed salt is preferably hydrochloride, compound 1 methyl ester sulfate, compound 1 ethyl ester hydrochloride, and compound 1 ethyl ester sulfate.
The pharmaceutical composition according to any one of claims 1 to 3, wherein the neuraminidase inhibitor is Oseltamivir (Oseltamivir) or its derivatives, zanamivir (zanamivir) or its derivatives , Peramivir (Peramivir) or a mixture of one or more of its derivatives in any ratio; the derivatives of oseltamivir, zanamivir, and peramivir are independently selected from The acid and/or base, or the ester, amide of the compound, or the carboxylic acid and salt thereof obtained after hydrolysis.
The pharmaceutical composition according to any one of claims 1 to 4, wherein the second active ingredient is oseltamivir free state and/or oseltamivir phosphate and/or oseltamivir carboxylic acid and/ Or oseltamivir carboxylate.
The pharmaceutical composition according to any one of claims 1 to 5, wherein the mass ratio of the first active ingredient to the second active ingredient is 100-600:75.
The pharmaceutical composition according to any one of claims 1 to 6, wherein the mass ratio of the first active ingredient to the second active ingredient is about 100:75, 200:75, 300:75, 400:75, 500:75, 600:75.
The pharmaceutical composition according to claim 1, wherein the first active ingredient is compound 1 and/or the sodium salt of compound 1 and/or the potassium salt of compound 1 and/or the hydrochloride of compound 1 and/or The calcium salt of compound 1 and/or the p-toluenesulfonate salt of compound 1; the second active ingredient is oseltamivir phosphate; the mass ratio of compound 1 to oseltamivir phosphate is 100-600:75.
The pharmaceutical composition according to claim 1, wherein the first active ingredient is compound 1 and/or the sodium salt of compound 1 and/or the potassium salt of compound 1 and/or the hydrochloride salt of compound 1 and/or The calcium salt of compound 1 and/or the p-toluenesulfonate salt of compound 1, the second active ingredient is oseltamivir phosphate, and the mass ratio of compound 1 to oseltamivir phosphate is 100:75,200:75,300:75,400: 75,500:75,600:75.
A preparation, characterized in that the preparation comprises the pharmaceutical composition according to any one of claims 1-9 and a pharmaceutically acceptable carrier.
The preparation according to claim 10, wherein the pharmaceutically acceptable carrier is selected from any one of fillers, binders, disintegrants, glidants, lubricants, and flavoring agents. Two or more.
The formulation according to claim 11, wherein the filler is selected from calcium carbonate, magnesium carbonate, calcium phosphate, calcium sulfate, magnesium oxide, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, sucrose, Any one or two or more of lactose, fructose, xylitol, mannitol, starch or its derivatives, dextrin, microcrystalline cellulose, etc. are mixed in any ratio to obtain a mixture;

The binder is selected from gum arabic, gelatin, tragacanth, dextrin, polyvinylpyrrolidone, starch and its derivatives, sodium alginate, sorbitol, syrup, hydroxypropyl methylcellulose, methyl cellulose , Hydroxypropyl cellulose, hydroxyethyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose and polymethacrylate any one or two or more of any Proportional mixing of the resulting mixture;

The disintegrant is selected from starch, alginic acid, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, croscarmellose sodium, cross-linked polyvinylpyrrolidone, low-substituted hydroxypropyl methylcellulose , A mixture obtained by mixing any one or more of microcrystalline cellulose and methyl cellulose in any ratio;

The glidant is a mixture obtained by mixing any one or two or more of colloidal silicon dioxide, powdered cellulose, magnesium trisilicate, silicon dioxide, and talc in any ratio;

The lubricant is selected from calcium stearate, glyceryl monostearate, glyceryl palmitate, magnesium stearate, microcrystalline cellulose, sodium benzoate, sodium chloride, sodium lauryl sulfate, hard A mixture obtained by mixing any one or two or more of sodium fat-based fumarate, talc, zinc stearate and polyethylene glycol in any ratio;

The flavoring agent is selected from any one or two or more of stevioside, aspartame and other flavors and sweeteners commonly used in the art in an arbitrary ratio.
The preparation according to any one of claims 10-12, wherein the preparation is an oral preparation; preferably a powder, granule, pellet, capsule, tablet, solution or lozenge.