WO2021175173A1 - Pharmaceutical composition for treating influenza and formulation containing the pharmaceutical composition - Google Patents

Abstract

Provided is a pharmaceutical composition. The pharmaceutical composition comprises a first active ingredient, a second active ingredient, and optionally a third active ingredient, and each active ingredient has a good drug synergy with each other. After a comprehensive evaluation, it is considered that the pharmaceutical composition has a good prospect of druggability.

Description

Medicinal composition for treating influenza and preparation containing the medicinal composition Technical field

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

Background technique

Influenza virus, or 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 protocol for influenza includes vaccination and antiviral drugs for chemotherapy and chemoprevention. Patent WO2018041263 discloses a series of pyrimidine derivatives with PB2Cap binding inhibitory effect. In vitro activity data showed that some compounds showed positive effects in the test of inhibiting influenza virus replication. In further animal tests, some compounds also showed significant therapeutic effects on influenza A virus H1N1 mouse infection model. Among them, compound 1 ( In Example 4, the comprehensive performance of WX-216) is relatively outstanding; in comparison with the main varieties that have been marketed in the same therapeutic field, compound 1 also shows a clear advantage, specifically, it is VX- in vitro against influenza A virus strains. It is about 10 times that of 787, which is 10 to tens of thousands times that of oseltamivir phosphate. It also shows a clear advantage over baloxavir dipivoxil and oseltamivir phosphate in the evaluation of drug resistance; therefore, compound 1 It is considered to have a good prospect of being a medicine.

Compound 1

Neuraminidase inhibitors are one of the important antiviral drugs, such as oseltamivir (OSE), zanamivir (zanamivir), peramivir (Peramivir), etc., such antiviral drugs It has obvious effect on influenza A virus and is a commonly used medicine for clinical treatment of viral influenza. 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.

Baloxavir marboxil, CAS: 1985606-14-1, developed by Shionoyoshi Co., Ltd., is a compound with cap-dependent endonuclease inhibition; Baloxavir marboxil enters the body After hydrolysis, it is metabolized into baloxavir, which has a good inhibitory effect on influenza A and B viruses and influenza virus strains resistant to oseltamivir; baloxavir dipivoxil preparation products were launched in Japan in 2018 It was first marketed under the trade name XOFLUZA, which is clinically used for the treatment of influenza A and influenza B.

According to in vitro drug resistance is a difficult problem that antiviral products need to face and overcome. Frederick G. Hayden et al, N Engl J Med, 2018, 379: 913-923 discloses the treatment data of baloxavir dipivoxil during phase II and phase III clinical trials. The data shows the performance of the baloxavir dipivoxil treatment group It has a significant therapeutic advantage over the oseltamivir group and the blank group; however, during the phase II and phase III clinical phases, 2.2% and 9.7% of the baloxavir dipivoxil group still had the treatment effect caused by the virus mutation, respectively A decline indicates that the virus has developed drug resistance. 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%.

It can be seen that influenza virus has the characteristics of high mutation rate and multiple inter-virus recombination phenomena, which affect the therapeutic effect of the drug; 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 exist It is unable to effectively inhibit/reduce the concentration of the virus in the body and cure the defects of the virus in a short time. The corresponding increase in the dosage and the lengthening of the treatment time also bring a greater risk of medication to the patient.

Combination medication is one of the effective solutions to the above-mentioned problems. Patent WO2017104691 discloses a compound preparation containing baloxavir dipivoxil. Compounds with RNA polymerase inhibition, compounds with M2 protein inhibition, compounds with PB2Cap binding inhibition, compounds with HA maturation inhibition, recombinant sialidase, compounds with reassembly inhibition, RNA interference Compounds, compounds having hemagglutinin receptor binding inhibitory effects, compounds having membrane fusion inhibitory effects of HA, compounds having nuclear translocation inhibitory effects of NP, compounds having CXCR inhibitory effects, and compounds having CRM1 inhibitory effects ; The compound with PB2Cap binding inhibitory effect can be VX-787 (Pimodivir). The activity data shows that the CI value of baloxavir dipivoxil in combination with VX-787 is 0.61, suggesting that baloxavir dipivoxil in combination with VX-787 It has a synergistic effect. In addition, Finberg RW et al., J Infect Dis, 2019 Mar 15; 219(7):1026-1034 also disclosed a combination experiment of VX-787 (Pimodivir) and oseltamivir, and the results showed that 600mg Pimodivir and Pimodivir The combined use of 75mg oseltamivir can minimize the load of influenza A virus. Regrettably, according to the information released in September 2020, based on the interim analysis data of the Phase III trial of hospitalized influenza A patients, Janssen Pharmaceuticals Inc. has stopped the clinical development of Pimodivir.

It can be seen that finding new solutions to prolong the service life of anti-influenza drugs with clinical application prospects to cope with the huge unmet clinical demand for anti-influenza virus drugs is a technical problem that the existing technology urgently needs to solve.

Summary of the invention

The first purpose of the present invention is to overcome the shortcomings of the prior art and provide a pharmaceutical composition for the treatment of influenza. The pharmaceutical composition utilizes the synergy between the active ingredients to solve the technology of influenza virus resistance. Problems, and can effectively reduce the risk of clinical medication, after comprehensive evaluation, it is believed that the pharmaceutical composition has a better prospect of preparing a medicine.

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

A pharmaceutical composition, wherein the pharmaceutical composition comprises a first active ingredient and a second active ingredient, wherein the first active ingredient is compound 1 or its corresponding ester or its corresponding salt, and the second active ingredient is Balo For savir dipivoxil and/or baloxavir, the mass ratio of the first active ingredient to the second active ingredient is about 1-50:1.

The first active ingredient refers to compound 1 or its corresponding ester or its corresponding salt, and specifically can be one of compound 1, the corresponding ester of compound 1, the salt of compound 1, and the salt of compound 1 corresponding to ester, or A mixture of two or more in any ratio, the aforementioned compound 1, the corresponding ester of compound 1, the salt of compound 1, and the salt of the corresponding ester of compound 1, each include its subordinate anhydrate, ansolvate, hydrate and solvate. The corresponding ester of compound 1 refers to the ester formed by compound 1 and organic acid, and preferred exemplary embodiments include but are not limited to the methyl, ethyl, propyl, isopropyl, n-butyl of compound 1 , Tert-butyl ester, etc.; the salt of compound 1 is a salt formed by compound 1 and an organic acid and/or an inorganic acid, or a salt formed by compound 1 and an organic base and/or an inorganic base. Preferred exemplary embodiments include But not limited to the hydrochloride, hydrobromide, sulfate, phosphate, benzenesulfonate, p-toluenesulfonate, methanesulfonate, tartrate, camphorsulfonate, lithium salt, sodium salt of compound 1 , Potassium salt, calcium salt, magnesium salt, aluminum salt, ammonia salt, ethylenediamine salt, triethylamine salt, etc.; the corresponding ester salt of compound 1 refers to the ester formed by compound 1 and organic acid and organic base and/ Or salts formed by inorganic bases and/or organic acids and/or inorganic acids. Preferred exemplary embodiments include, but are not limited to, the hydrochloride of compound 1 methyl ester, the sulfate of compound 1 methyl ester, and the salt of compound 1 ethyl ester. Hydrochloride, compound 1 ethyl sulfate, etc. And, the aforementioned 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 lower anhydrate, ansolvate, hydrate, and solvate thereof.

The second active ingredient refers to baloxavir dipivoxil and/or baloxavir, and also includes its subordinate anhydrates, ansolvates, hydrates and solvates. For example, baloxavir dipivoxil may be Anhydrous baloxavir dipivoxil may also be a hydrate of baloxavir dipivoxil or a solvate of baloxavir dipivoxil.

According to a preferred embodiment of the present invention, the mass ratio of the first effective ingredient to the second effective ingredient is about 1-50:1; further, the mass ratio of the first effective ingredient to the second effective ingredient About 5-40:1; furthermore, the mass ratio of the first effective ingredient to the second active ingredient is about 5-20:1; in particular, the mass of the first effective ingredient and the second effective ingredient The ratio may be approximately 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1.

Unless otherwise specified, the quality of the salt mentioned in the present invention refers to the mass in terms of free base/acid equivalent, and the hydrate/solvate refers to the mass in terms of dry matter (on an equivalent basis). anhydrous basis).

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is compound 1, and the second active ingredient is anhydrous baloxavir dipivoxil; in the aforementioned pharmaceutical composition, compound 1 and anhydrous baloxavir The mass ratio of the ester is about 1-50:1; further, the mass ratio of the compound 1 and the anhydrous baloxavir dipivoxil is about 5-40:1; and further, the compound 1 and the anhydrous baloxavir dipivoxil are about 5-40:1. The mass ratio of loxavir dipivoxil is about 5-20:1. Specifically, the mass ratio of the first effective ingredient to the second effective ingredient may be about 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35: 1,40:1,45:1,50:1.

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 anhydrous baloxavir dipivoxil; the sodium salt of compound 1 in the aforementioned pharmaceutical composition is combined with The mass ratio of baloxavir dipivoxil anhydrate is about 1-50:1; further, the mass ratio of the sodium salt of compound 1 to baloxavir dipivoxil anhydrate is about 5-40:1; Further, the mass ratio of the sodium salt of compound 1 to baloxavir dipivoxil anhydrate is about 5-20:1. Specifically, the mass ratio of the sodium salt of compound 1 to anhydrous baloxavir dipivoxil may be about 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30: 1,35:1,40:1,45:1,50:1.

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is the hydrochloride of compound 1, and the second active ingredient is anhydrous baloxavir dipivoxil; the hydrochloric acid of compound 1 in the aforementioned pharmaceutical composition The mass ratio of salt to anhydrous baloxavir dipivoxil is about 1-50:1; further, the mass ratio of the hydrochloride salt of compound 1 to anhydrous baloxavir dipivoxil is about 5-40:1; Furthermore, the mass ratio of the hydrochloride salt of compound 1 to anhydrous baloxavir dipivoxil is about 5-20:1. Specifically, the mass ratio of the hydrochloride salt of compound 1 to anhydrous baloxavir dipivoxil may be about 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30 :1,35:1,40:1,45:1,50:1.

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 anhydrous baloxavir dipivoxil; the potassium salt of compound 1 in the aforementioned pharmaceutical composition is combined with The mass ratio of anhydrous baloxavir dipivoxil is about 1-50:1; further, the mass ratio of the potassium salt of compound 1 to anhydrous baloxavir dipivoxil is about 5-40:1; and further The mass ratio of the potassium salt of compound 1 to anhydrous baloxavir dipivoxil is about 5-20:1. Specifically, the mass ratio of the potassium salt of compound 1 to anhydrous baloxavir dipivoxil may be about 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30: 1,35:1,40:1,45:1,50:1.

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 anhydrous baloxavir dipivoxil; the calcium salt of compound 1 in the aforementioned pharmaceutical composition is combined with The mass ratio of anhydrous baloxavir dipivoxil is about 1-50:1; further, the mass ratio of the calcium salt of compound 1 to anhydrous baloxavir dipivoxil is about 5-40:1; and further The mass ratio of the calcium salt of compound 1 to anhydrous baloxavir dipivoxil is about 5-20:1. Specifically, the mass ratio of the calcium salt of compound 1 to anhydrous baloxavir dipivoxil may be about 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30: 1,35:1,40:1,45:1,50:1.

In a preferred technical solution of the present invention, the first active ingredient in the pharmaceutical composition is the benzoate of compound 1, and the second active ingredient is anhydrous baloxavir dipivoxil; the benzene of compound 1 in the aforementioned pharmaceutical composition The mass ratio of formate to anhydrous baloxavir dipivoxil is about 1-50:1; further, the mass ratio of the benzoate of compound 1 to anhydrous baloxavir dipivoxil is about 5-40 :1; Furthermore, the mass ratio of the benzoate of compound 1 to anhydrous baloxavir dipivoxil is about 5-20:1. Specifically, the mass ratio of the benzoate of compound 1 to anhydrous baloxavir dipivoxil may be about 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1,35:1,40:1,45:1,50:1.

The inventor’s surprising discovery in the experiment: the aforementioned drug containing the first active ingredient (compound 1 or its corresponding ester or its corresponding salt) and the second active ingredient (baloxavir dipivoxil or baloxavir) The composition, when the mass ratio of the two active ingredients is in a specific range, has a better drug synergistic effect, which is manifested as a significantly better anti-influenza virus effect than a single ingredient, and also exhibits a higher mass ratio than the aforementioned specific range. The pharmaceutical composition has significantly better anti-influenza virus effect. It can be seen 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 a better anti-influenza virus effect; the rest of the drug properties are integrated (such as mixture stability, fluidity, etc.) After the evaluation, it is believed that the pharmaceutical composition has a better prospect of preparing medicine.

Further, the aforementioned pharmaceutical composition in the first object of the present invention, on the basis of including the first effective ingredient and the second effective ingredient, further includes a third effective ingredient, and the third effective ingredient is neuraminic acid. Enzyme inhibitors; the definitions and corresponding ranges of the first active ingredient and the second active ingredient are the same as the aforementioned pharmaceutical composition in the first object of the present invention.

Examples of the neuraminidase inhibitor referred to by the third active ingredient preferably include oseltamivir (Oseltamivir) or its derivatives, zanamivir (zanamivir) or its derivatives, peramivir (Peramivir) Or a mixture of one or two or more of the derivatives thereof in any ratio; the derivatives of oseltamivir, zanamivir, and peramivir are independently selected from The acid and/or base may also be salts, esters, amides, etc., which are further formed by the compound, or other derivatives such as carboxylic acids and their salts obtained after hydrolysis, and each includes its subordinate anhydrate, ansolvate, Hydrates and solvates, such as oseltamivir or its derivatives can be selected as oseltamivir free state, or oseltamivir phosphate, oseltamivir carboxylic acid, oseltamivir carboxylate Wait.

Specifically, in the pharmaceutical composition containing the third active ingredient, as mentioned above, the first active ingredient is compound 1 or its corresponding ester or its corresponding salt, and the second active ingredient is baloxavir dipivoxil And/or baloxavir; the mass ratio of the first active ingredient, the second active ingredient and the third active ingredient in the pharmaceutical composition is about 1-50:1:20-100; preferably, the pharmaceutical composition The mass ratio of the first effective ingredient, the second effective ingredient and the third effective ingredient is about 5-40:1:1:30-80; preferably, the first effective ingredient, the second effective ingredient and the first effective ingredient in the pharmaceutical composition The mass ratio of the three active ingredients is about 5-20:1: 1:30-80. Specifically, the mass ratio of the first active ingredient, the second active ingredient and the third active ingredient in the pharmaceutical composition may be about 1: 1:33,5:1:33,10:1:33,15:1:33,20:1:33,25:1:33,30:1:33,35:1:33,40:1: 33,45:1:33,50:1:33.

In a preferred embodiment of the present invention, the aforementioned pharmaceutical composition containing the first active ingredient, the second active ingredient and the third active ingredient is a combination of compound 1, anhydrous baloxavir dipivoxil, and oseltamivir phosphate.

In a preferred embodiment of the present invention, the aforementioned pharmaceutical composition containing the first active ingredient, the second active ingredient and the third active ingredient is the sodium salt of compound 1, anhydrous baloxavir dipivoxil, and oseltamivir phosphate The combination.

In a preferred embodiment of the present invention, the aforementioned pharmaceutical composition containing the first active ingredient, the second active ingredient and the third active ingredient is the potassium salt of compound 1, anhydrous baloxavir dipivoxil, and oseltamivir phosphate The combination.

In a preferred embodiment of the present invention, the aforementioned pharmaceutical composition containing the first active ingredient, the second active ingredient and the third active ingredient is the calcium salt of compound 1, anhydrous baloxavir dipivoxil, and oseltamivir phosphate The combination.

In a preferred embodiment of the present invention, the aforementioned pharmaceutical composition containing the first active ingredient, the second active ingredient and the third active ingredient is the hydrochloride salt of compound 1, anhydrous baloxavir dipivoxil, and oseltamivir phosphate The combination of salt.

In a preferred embodiment of the present invention, the aforementioned pharmaceutical composition containing the first active ingredient, the second active ingredient and the third active ingredient is the benzoate of compound 1, anhydrous baloxavir dipivoxil, and oseltamivir The combination of phosphates.

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 is prepared by a conventional mixing method in the field. More specifically, the preparation method may be a direct mixing method, an equal incremental method, etc., and the mixing equipment used may be common in the field depending on the preparation scale. The mixing 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 pharmaceutically acceptable excipients. The pharmaceutically acceptable excipients particularly refer to pharmaceutically acceptable carriers, which can be specifically selected from fillers, binders, and disintegrants. Any one or two or more of additives, glidants, lubricants, and flavoring agents.

Specifically, the filler is selected from calcium carbonate, magnesium carbonate, calcium phosphate, calcium sulfate, magnesium oxide, sucrose, lactose, fructose, xylitol, mannitol, starch or its derivatives, dextrin, microcrystalline cellulose Any one or a mixture of two or more in any ratio.

Specifically, the binder is selected from gum arabic, gelatin, tragacanth, dextrin, polyvinylpyrrolidone, sodium alginate, sorbitol, syrup, hydroxypropyl methyl cellulose, methyl cellulose, hydroxypropyl Any one or two or more in any ratio of base cellulose, hydroxyethyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose and polymethacrylate, etc. mixture.

Specifically, the disintegrant is selected from any one or two or more in any ratio of alginic acid, croscarmellose sodium, cross-linked polyvinylpyrrolidone and low-substituted hydroxypropyl methylcellulose, etc. mixture.

Specifically, the glidant is selected from any one or a mixture of two or more in any ratio among powdered cellulose, magnesium trisilicate, silicon dioxide, and talc.

Specifically, the lubricant is selected from calcium stearate, glyceryl monostearate, glyceryl palmitate, magnesium stearate, sodium benzoate, sodium chloride, sodium lauryl sulfate, magnesium stearate , Sodium stearyl fumarate, zinc stearate, polyethylene glycol, etc. Any one or a mixture of two or more in any 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, or a mixture of two or more in any ratio.

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

In the aforementioned formulation, the specification of Compound 1 or its corresponding ester or its corresponding salt in the said formulation, that is, the amount of Compound 1 or its corresponding ester or its corresponding salt contained in a unit preparation can be 50-1500 mg; Further, the specification of the preparation is 100-1000 mg; specifically, the specification of compound 1 or its corresponding ester or its corresponding salt in the preparation is 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg , 900mg, 1000mg.

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 quality ratios have significantly better comprehensive anti-influenza virus effects, and have a better market prospect.

The fourth object of the present invention is to provide a pharmaceutical application, that is, the pharmaceutical composition according to the first object of the present invention and/or the preparation according to the third object of the present invention is used in the preparation of drugs for the treatment of influenza. The purpose of the aspect.

The inventor surprisingly discovered that, based on the synergy between the drugs, the pharmaceutical composition of the present invention and the preparation containing the pharmaceutical composition show better anti-influenza therapeutic effects than when used alone, can effectively reduce the dosage, and It is beneficial to avoid the emergence of drug resistance; further, the influenza targeted for the pharmaceutical use is influenza A; and further, the influenza targeted for the pharmaceutical use is caused by the A/PR/8/34 influenza A virus Influenza A.

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 or its corresponding ester or its corresponding salt) and a second active ingredient (baloxavir dipivoxil) with a mass ratio in a specific range And/or baloxavir), and can optionally further contain a third active ingredient (neuraminidase inhibitor), each active ingredient has a better drug synergistic effect, and after comprehensive evaluation, it is considered to have a better finished drug prospect;

2. A preparation method of the pharmaceutical composition of the present invention 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 exhibits a significantly better anti-influenza virus effect than a preparation containing a single component, and also exhibits a better anti-influenza virus effect 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;

4. A pharmaceutical application is provided, which utilizes the synergy between drugs to achieve better therapeutic effects.

The contribution of the present invention is the discovery of synergy between drugs within a specific range. Therefore, those skilled in the art can understand that when the type and ratio of the effective ingredients in the unit dosing unit are within the range described in the present invention, it can be used. It is considered that the technical solution protected by the present invention is used; specifically, the aforementioned unit dosing unit refers to the smallest unit for clinical use, such as: unit tablet, unit capsule, unit bottle of oral liquid, unit package The granules and so on.

Detailed ways

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 baloxavir dipivoxil and baloxavir of the present invention can be prepared by referring to the methods disclosed in the prior art, for example, referring to the methods disclosed in Synthesis Example 1 and Synthesis Example 2 of WO2017104691. Specifically, the baloxavir dipivoxil used in the examples of the present invention are all self-made bulk drugs with a purity of >99%; the baloxavir used in the examples of the present invention are all self-made bulk drugs with a purity of >99%.

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%.

Example 1 Preparation of compound 1

The free substance of Compound 1 was prepared by the method disclosed in Example 4 of WO2018041263, and the structure characterization proved that the obtained product was the free substance of 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 hydrochloric acid (0.98mL, dissolved in 9mL THF), stir for 12 hours at 30°C, filter the solid, and dry the filter cake at 40°C under vacuum to obtain compound 1 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 (0.101 g) was added to acetone (2 mL) and slurried for 12 h to obtain the p-toluenesulfonate salt 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 solid, and filter cake at 40°C It was dried in vacuum to obtain the potassium salt of compound 1.

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 Dry under vacuum at °C 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. The solid After filtration, the filter cake was vacuum dried at 40° C. to obtain the calcium salt of compound 1.

Example 7

Take 200 g of the compound 1 free substance prepared in Example 1, and mix it with 20 g of baloxavir dipivoxil uniformly in an equal increment method to obtain a pharmaceutical composition with a mass ratio of 10:1.

Example 8

Using the same method as in Example 7, the mass ratios of compound 1 free substance and baloxavir dipivoxil were prepared respectively as 1:1, 5:1, 10:1, 15:1, 25:1, 30:1, 35 :1,40:1,45:1,50:1 pharmaceutical composition.

Using the same method as in Example 7, using the different salts of Compound 1 prepared in Examples 2-6 to replace the free substance of Compound 1, they can be prepared with Baloxavir Dipivoxil in a mass ratio of 1:1, 5:1. ,10:1,15:1,20:1,25:1,30:1,35:1,40:1,45:1,50:1 pharmaceutical composition, the quality of the salt is referred to The quality of the free substance meter (free base/acid equivalent).

Example 9

In vitro activity experiment, to investigate the combination of compound 1 and the active metabolite baloxavir. The pharmaceutical composition of compound 1 and baloxavir in the mass ratio described in Example 8 was prepared, influenza virus strains were selected, and the antiviral effects of each group were tested respectively.

In vitro activity experiments show that compound 1 and baloxavir have a certain synergistic effect in vitro, and have better anti-influenza effects than the single use of compound 1 and the single use of baloxavir experimental group.

Example 10

In vivo efficacy study of the drug against A/PR/8/34 influenza A virus {A/PR/8/34(H1N1)}.

Study on the protection of drugs against A/PR/8/34 influenza A virus death (administration 2h after A/PR/8/34 A virus infection)

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

Administration group for 5 consecutive days: 2 hours after infection, intragastric administration, twice a day, for 5 consecutive days, the drug group is set as follows:

The first group: Compound 1: 1.5mg/kg;

The second group: Compound 1: 5mg/kg;

The third group: Compound 1: 20mg/kg;

The fourth group: Baloxavir Dipivoxil: 0.3mg/kg;

The fifth group: Oseltamivir phosphate 10mg/kg;

The sixth group: compound 1 + baloxavir dipivoxil = 0.1 mg/kg + 0.3 mg/kg;

The seventh group: compound 1 + baloxavir dipivoxil = 0.3 mg/kg + 0.3 mg/kg;

The eighth group: compound 1 + baloxavir dipivoxil = 1.5 mg/kg + 0.3 mg/kg;

The ninth group: compound 1+baloxavir dipivoxil=5mg/kg+0.3mg/kg;

The tenth group: compound 1+baloxavir dipivoxil=20mg/kg+0.3mg/kg;

The eleventh group: compound 1+ baloxavir dipivoxil+ oseltamivir phosphate=1.5mg/kg+0.3mg/kg+10mg/kg; and set up a blank control group and a virus control group, 9 in each group; Observe from the first day of virus infection and continue to 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 (%) = the 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 showed a significant dose-dependent effect in reducing mortality, and the mortality of the 1.5 mg/kg group (first group), 5 mg/kg group (second group), and 20 mg/kg group (third group) were respectively 88.89, 44.44 and 0%;

Baloxavir Dipivoxil: The mortality rate of mice in the 0.3 mg/kg administration group (group 4) was 77.78%, and the mortality rate of mice in the oseltamivir phosphate: the 10 mg/kg administration group (group 5) was 100. %; compound 1 + baloxavir dipivoxil = 0.1 mg/kg + 0.3 mg/kg (group 6), the mortality rate of mice is 77.88%, which does not show sufficient advantages compared with the single prescription;

Compound 1 + Baloxavir Dipivoxil = 0.3 mg/kg + 0.3 mg/kg (group 7) The mortality rate of mice was 33.33%, Compound 1 + Baloxavir Dipivoxil = 1.5 mg/kg + 0.3 mg/kg ( The eighth group) mouse mortality rate was 11.11%, compound 1+ baloxavir dipivoxil = 5mg/kg+0.3mg/kg (the ninth group) mouse mortality rate was 0%, and the treatment effect was better than the single prescription (the fourth group). ) Significantly increased, and compound 1 + baloxavir dipivoxil + oseltamivir phosphate = 1.5 mg/kg + 0.3 mg/kg + 10 mg/kg (the tenth group) mouse mortality rate was 0%;

Compound 1 + Baloxavir Dipivoxil = 20 mg/kg + 0.3 mg/kg (group ten) The mortality rate of mice was also 0%, the same as the third group, but some of the surviving mice had lower mental states than those of the sixth and seventh groups. In the eight groups of surviving mice, it can be seen that under the premise that the therapeutic effect cannot be further improved, there may be potential drug risks due to excessive use of drugs;

It can be seen that after compound 1 was used in combination with 0.3 mg/kg baloxavir dipivoxil at doses of 0.3 mg/kg, 1.5 mg/kg and 5 mg/kg, the mortality rate was significantly reduced, and the synergistic effect was obvious. The specific results are shown in Table 1 below. Show:

Table 1 Research on the efficacy of anti-influenza in vivo

In vivo activity experiments show that compound 1 and baloxavir dipivoxil have a certain synergistic effect. When the ratio of compound 1 and baloxavir dipivoxil is 1:1, 5:1, and 16.7:1, it is better than using compound 1 alone. And single use of baloxavir dipivoxil has better anti-influenza virus effect.

Comprehensive in vivo efficacy study results show that when compound 1 or its corresponding ester or its corresponding salt, and baloxavir dipivoxil or baloxavir have a ratio of 1 to 50:1, there is a certain degree of difference between the active ingredients Drug synergistic effect; when the ratio of compound 1 or its corresponding ester or its corresponding salt to baloxavir dipivoxil or baloxavir is 5-40:1, the active ingredients have a better drug synergistic effect; When the ratio of compound 1 or its corresponding ester or its corresponding salt to baloxavir dipivoxil or baloxavir is 5-20:1, the active ingredients have a good drug synergistic effect; the aforementioned synergy performance In order to have a significantly better anti-influenza virus effect than a single component, it also exhibits a significantly better anti-influenza virus effect than a pharmaceutical composition with a mass ratio outside the aforementioned specific range. Specifically, compound 1 or its corresponding ester or its corresponding salt, and the mass ratio of baloxavir or baloxavir of 1:1, 5:1 and 16.7:1 have significantly better anti-influenza virus effect The effect of using compound 1 or its corresponding ester or its corresponding salt alone, and the effect of using baloxavir dipivoxil or baloxavir alone, the comprehensive therapeutic effect is also significantly better than that of a pharmaceutical composition whose quality ratio is not within the specific range. Anti-flu virus effect.

In addition, the experimental data also showed that the combined use of compound 1, baloxavir dipivoxil and oseltamivir phosphate can effectively improve the survival rate of mice, showing that compound 1 or its corresponding ester or its corresponding salt, baloxavir Ester or baloxavir, oseltamivir or its corresponding salt have obvious drug synergistic effect; when compound 1 or its corresponding ester or its corresponding salt, and the second active ingredient is baloxavir dipivoxil And/or when the mass ratio of baloxavir and the third active ingredient oseltamivir or its corresponding salt is about 1-50:1:20-100, the pharmaceutical composition has a good anti-influenza virus effect; When the mass ratio of the first effective ingredient, the second effective ingredient and the third effective ingredient in the pharmaceutical composition is about 5-40:1:1:30-80, the pharmaceutical composition has a better anti-influenza virus effect; and When the mass ratio of the first effective ingredient, the second effective ingredient and the third effective ingredient in the pharmaceutical composition is about 5-20:1:1:30-80, the anti-influenza virus effect of the pharmaceutical composition is optimal.

Example 11

The pharmaceutical composition obtained in Example 7 was mixed with an appropriate amount of fillers, binders, and disintegrants and then directly compressed to prepare tablets (200 mg+20 mg).

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 (30)

1. 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 or its corresponding ester or its corresponding salt, and the second active ingredient is Baloxavir Dipivoxil and/or Baloxavir, the mass ratio of the first active ingredient to the second active ingredient is 1-50:1,

   
2. The pharmaceutical composition according to claim 1, wherein the first active ingredient is compound 1 or its corresponding ester or its corresponding salt is compound 1, the corresponding ester of compound 1, the salt of compound 1, and the corresponding ester of compound 1. A mixture of one or two or more of the salts in any ratio.
3. The pharmaceutical composition according to claim 2, wherein the ester corresponding to compound 1 refers to the methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl of compound 1; the salt of compound 1 It is the hydrochloride, hydrobromide, sulfate, phosphate, benzenesulfonate, p-toluenesulfonate, methanesulfonate, tartrate, camphorsulfonate, lithium salt, sodium salt, potassium of compound 1 Salt, calcium salt, magnesium salt, aluminum salt, ammonia salt, ethylenediamine salt, triethylamine salt; the corresponding ester salt of compound 1 is the hydrochloride salt of compound 1 methyl ester, the sulfate salt of compound 1 methyl ester, and the compound 1 Ethyl ester hydrochloride, compound 1 Ethyl sulfate salt.
4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the second active ingredient refers to baloxavir and/or baloxavir refers to baloxavir, baloxavir, A mixture of Baloxavir Dipivoxil and Baloxavir in any ratio.
5. The pharmaceutical composition according to any one of claims 1-4, wherein the second active ingredient is baloxavir dipivoxil.
6. 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 5-40:1.
7. 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 5-20:1.
8. The pharmaceutical composition according to claim 1, wherein the mass ratio of the first active ingredient to the second active ingredient is 1:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1.
9. The pharmaceutical composition according to claim 1, wherein the first active ingredient in the pharmaceutical composition 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 baloxavir dipivoxil, and the mass ratio of compound 1 to baloxavir dipivoxil is 5-40:1.
10. The pharmaceutical composition according to claim 1, wherein the first active ingredient in the pharmaceutical composition 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 baloxavir dipivoxil, and the mass ratio of compound 1 to baloxavir dipivoxil is 5-20:1.
11. The pharmaceutical composition according to claim 1, wherein the first active ingredient in the pharmaceutical composition 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 baloxavir dipivoxil, and the mass ratio of the first active ingredient to baloxavir dipivoxil is 1:1, 5: 1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1.
12. The pharmaceutical composition according to any one of claims 1-11, wherein the pharmaceutical composition further comprises a third active ingredient, and the third active ingredient is a neuraminidase inhibitor.
13. The pharmaceutical composition according to claim 12, wherein the third active ingredient is one or both of oseltamivir or its derivatives, zanamivir or its derivatives, peramivir or its derivatives A mixture of more than one kind in any ratio.
14. The pharmaceutical composition according to claim 12 or 13, wherein the third active ingredient is oseltamivir phosphate.
15. The pharmaceutical composition according to any one of claims 12-14, wherein the mass ratio of the first active ingredient, the second active ingredient and the third active ingredient is 1-50:1:20-100.
16. The pharmaceutical composition according to any one of claims 12-14, wherein the mass ratio of the first active ingredient, the second active ingredient and the third active ingredient is 5-40:1:30-80.
17. The pharmaceutical composition according to any one of claims 12-14, wherein the mass ratio of the first active ingredient, the second active ingredient and the third active ingredient is 5-20:1:30-80.
18. The pharmaceutical composition according to any one of claims 12-14, wherein the mass ratio of the first active ingredient, the second active ingredient and the third active ingredient is 1:1:33, 5:1:33, 10: 1:33,15:1:33,20:1:33,25:1:33,30:1:33,35:1:33,40:1:33,45:1:33,50:1: 33.
19. A preparation, characterized in that the preparation comprises the pharmaceutical composition according to any one of claims 1-18 and pharmaceutically acceptable excipients.
20. The formulation according to claim 19, wherein the pharmaceutically acceptable excipients are selected from any one of fillers, binders, disintegrants, glidants, lubricants, and flavoring agents. Two or more.
21. The preparation according to claim 20, wherein the filler is selected from calcium carbonate, magnesium carbonate, calcium phosphate, calcium sulfate, magnesium oxide, sucrose, lactose, fructose, xylitol, mannitol, starch and Any one or a mixture of two or more of derivatives, dextrin, and microcrystalline cellulose in any ratio.
22. The preparation according to claim 20, wherein the binder is selected from gum arabic, gelatin, tragacanth, dextrin, polyvinylpyrrolidone, sodium alginate, sorbitol, syrup, hydroxypropyl methyl Any one of cellulose, methyl cellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, ethyl cellulose, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose and polymethacrylate A mixture of two or more in any ratio.
23. The preparation according to claim 20, wherein the disintegrant is selected from the group consisting of alginic acid, croscarmellose sodium, cross-linked polyvinylpyrrolidone and low-substituted hydroxypropyl methylcellulose. One or two or more mixtures in any ratio.
24. The formulation according to claim 20, wherein the glidant is selected from any one or two or more selected from powdered cellulose, magnesium trisilicate, silicon dioxide, and talc in any ratio. The resulting mixture.
25. The preparation of claim 20, wherein the lubricant is selected from calcium stearate, glyceryl monostearate, glyceryl palm stearate, magnesium stearate, sodium benzoate, sodium chloride, Any one or a mixture of two or more of sodium lauryl sulfate, magnesium stearate, sodium stearyl fumarate, zinc stearate and polyethylene glycol in any ratio.
26. The preparation according to claim 20, wherein the corrective agent is selected from any one or more of stevioside, aspartame, and other flavors and sweeteners commonly used in the art. Proportion of the mixture.
27. The preparation according to any one of claims 19-26, wherein the preparation is an oral preparation, and the oral preparation is selected from the group consisting of powders, granules, pellets, capsules, tablets, solutions, or lozenges.
28. The use of the pharmaceutical composition according to any one of claims 1-18 or the preparation according to any one of claims 19-27 in the preparation of a medicament for the treatment of influenza.
29. The application according to claim 28, wherein the influenza is influenza A.
30. The application according to any one of claims 28-29, wherein the influenza is influenza A caused by A/PR/8/34 A virus.