RU2842252C1 - Solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid containing a zwitterion dimer of formula (III), characterized by that the solid form relates to a crystalline form characterized by a monoclinic system of the crystal lattice and values of edges of the crystal cell (in angstrom, Å): 8.71±0.50, 20.01±0.50 and 12.16±0.50 and angle value (in degrees) 93.10 ± 5.00°. Invention relates to a method for producing a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid according to invention, comprising reacting histamine with glutaric anhydride and treatment of obtained product with mineral acid at pH 5.2-5.7 followed by alkalization or addition of base until precipitation of solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid, containing 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid in form of a dimer of formula (III). Invention also relates to a pharmaceutical composition for the treatment and/or prevention of diseases caused by viral infections, comprising a solid form of the invention and at least one pharmaceutically acceptable excipient, wherein the viral infections are a parainfluenza virus, an influenza virus, including influenza A, B, C and D.

EFFECT: solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid containing zwitterion dimer, used for treating and/or preventing diseases caused by viral infections, wherein the viral infections are a parainfluenza virus, an influenza virus, including influenza A, B, C and D, which also has anti-inflammatory and antipyretic action.

14 cl, 14 dwg, 5 tbl, 13 ex

Description

Field of technology

The invention relates to the fields of organic chemistry, pharmacology and the chemical-pharmaceutical industry, namely to new solid forms of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid of formula (I), containing 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid in the form of a zwitterion of formula (II) or a dimerazwitterion of formula (III). The invention also relates to a pharmaceutical composition and a medicinal product containing the said forms of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid. The invention can find application in the treatment and/or prevention of diseases caused by viral infections.

State of the art

Infectious diseases are the third leading cause of death worldwide, most of which are viral diseases. Among viral infections, herpes viruses are one of the leading causes due to their widespread distribution. According to WHO, about 90% of the world's population has manifestations of herpes infection, and mortality from these infections is second only to viral hepatitis. Therefore, the problem of herpes diseases is especially relevant today. Science knows more than 100 types of herpes virus, of which 8 antigenic varieties cause diseases in humans (herpes simplex viruses of types 1 and 2, chickenpox and shingles (Varicellazoster), cytomegalovirus, Epstein-Barr virus, human herpes viruses of types 6, 7 and 8). Herpes viruses can remain in the human body for life and cause diseases with various manifestations from asymptomatic carriage to severe forms with the development of herpes encephalitis, meningitis, ending in death. The localization of herpes rashes is different - skin, mucous membranes of the oral cavity and genitals, cornea, internal organs. For the treatment and prevention of relapses of herpes infection, there is a large selection of drugs with a pronounced antiviral effect, but these drugs have a number of side effects, including a decrease in the body's resistance to viruses (O.V. RUBLEVA, On the issue of therapy and prevention of viral infections, RUSSIAN MEDICAL JOURNAL, 2012, Vol. 20, 11, pp. 571-574).

Many viruses affect the respiratory tract, nasal passages, throat, bronchi, lungs, and therefore such viral infections are grouped into acute respiratory viral infections (ARVI), since they also have common clinical symptoms. Influenza viruses also belong to the ARVI group, but are allocated to a separate group due to the fact that they affect the entire body systemically. ARVI can be caused by more than 200 different viruses, representatives of 4 families of RNA-containing viruses (orthomyxoviruses, paramyxoviruses, coronaviruses, and picornaviruses) and 2 families of DNA-containing viruses (adenoviruses and herpesviruses). In terms of infection frequency, influenza accounts for about 15% (type A - 12%, B - 3%), parainfluenza and rhinoviruses up to 50%, adenovirus up to 5%, respiratory syncytial virus (RS) - 4%, mycoplasma - about 2%, enteroviruses - about 1%, mixed infections - about 23% of cases. The high-risk group includes children, in whom respiratory tract diseases account for up to 90% of all infectious pathology and 65% of all registered diseases (O.V. RUBLEVA, On the issue of therapy and prevention of viral infections, RUSSIAN MEDICAL JOURNAL, 2012, Vol. 20, 11, pp. 571-574; E.P. SELKOVA, Prevention and treatment of acute respiratory viral infections. Use of amixin, Handbook for doctors, Moscow, 2004).

All ARIs are characterized by symptoms of general intoxication (headache, weakness, muscle pain, loss of appetite), fever, catarrhal symptoms (nasal congestion, runny nose, cough, sore throat). The localization of the lesion largely depends on the type of pathogen. Thus, rhinovirus diseases are characterized by the predominance of rhinitis, adenovirus infections - nasopharyngitis, parainfluenza is characterized by pharyngitis, and flu - tracheitis, respiratory syncytial virus affects the bronchi. Other symptoms are also possible, for example, adenoviruses can cause conjunctivitis and keratitis. Usually, symptoms of ARI persist for 3-7 days (cough can be observed for a longer time), the possible duration of flu is 1-2 weeks. Complications include laryngitis, tonsillitis, pharyngitis and tracheitis of bacterial origin, various forms of sinusitis (sinusitis, ethmoiditis, frontal sinusitis), as well as otitis, eustachitis, and possibly bronchitis and pneumonia. The main causes of complications are immune system disorders leading to long-term immunodeficiencies combined with a sharp decrease in the body's antibacterial resistance. With the right tactics for treating ARVI, the risk of complications is significantly reduced (Yu.Z.GENDON, Etiology of acute respiratory diseases, VACCINATION, 2001, Vol. 5, 17, pp. 4-5).

The most effective way to control seasonal influenza is vaccination to date. There are influenza vaccines, which are divided into two classes depending on the manufacturing technology: live and inactivated. The main disadvantage of vaccination is that its effect is limited to influenza viruses only, and there is no protection against other pathogens of acute respiratory viral infections. A promising direction for prevention is the use of agents to activate the body's non-specific resistance (A.A. ZAITSEV, Directions of Pharmacotherapy and Prevention of Acute Respiratory Viral Infections, RUSSIAN MEDICAL JOURNAL, 2009, Vol. 17, 23, pp. 1525-1529). The prophylactic effectiveness of antiviral agents during an outbreak is high and reaches 70-80%. At the same time, prophylaxis with antiviral agents can be carried out both for immunized individuals and for individuals who have not been vaccinated.

Thus, there is a need to develop new stable agents and affordable therapeutic forms for the treatment and prevention of influenza and acute respiratory viral infections in order to create strategic state reserves in case of epidemics or pandemics.

The closest analogue for the invention under consideration is the compound 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid. It is known to be used as an antioxidant (RU 2141483 C1) and antiviral (EA 020283 B1, RU 2496512 C1) agent.

2-(Imidazol-4-yl)ethanamide of pentanedioic-1,5 acid has found application in clinical practice as a drug effective against influenza A virus (Registration Certificate LP-006482).

One of the urgent tasks in the development of drugs remains the production of an active substance that is stable over a long shelf life. Considering that it is impossible to predict the antigenic structure of a future epidemic (pandemic) virus, it is difficult to design effective vaccines in advance, so it is important to have drugs in the arsenal for the prevention and treatment of diseases caused by highly pathogenic strains of viruses. At the same time, as the situation with the COVID-19 pandemic around the world has shown (Y. WANG et al., Unique epidemiological and clinical features of the emerging 2019 novel coronavirus pneumonia (COVID-19) implicate special control measures, JOURNAL OF MEDICAL VIROLOGY, 2020, Vol. 92, No. 6, pp. 568-576), in order to timely provide citizens with antiviral drugs in a pandemic, it is advisable to create state reserves of such funds. Strategic state reserves in case of pandemics are subject to long-term storage and renewal upon the expiration of the reserved drugs. In such a situation, it is extremely important to increase long-term stability in order to extend shelf life and reduce government costs for the creation and renewal of strategic reserves of antiviral agents.

Among the drugs used for influenza and acute respiratory viral infections, priority is given to etiotropic drugs. One of these compounds is 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid. Accordingly, the development of more stable forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid suitable for long-term storage is one of the most pressing tasks in the current conditions.

In order to increase the shelf life of compositions containing 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid, it was previously proposed to use a substance additionally containing N,N'- bis-[2-(1H-imidazol-4-yl)ethyl]pentanediamide (RU 2746692 C1). The complexity of this solution is the need to have N,N'- bis-[2-(1H-imidazol-4-yl)ethyl]pentanediamide in the composition containing 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid. The present technical solution proposes a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid, the storage of which is possible without the addition of auxiliary substances.

Thus, the objectives of the present invention are to develop an industrially available, storage-stable form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid for the treatment and prevention of respiratory diseases, in particular influenza and acute respiratory viral infections, providing high antiviral activity, as well as to expand the arsenal of drugs with antiviral action and suitable for the creation of strategic state reserves in the event of epidemics or pandemics.

Disclosure of invention

Technical result

Compounds with charged groups (ammonium, phosphonium ions, etc.) attract the attention of medicinal chemists due to their unique physicochemical properties. For example, it is known that phosphonium salts accumulate in inflamed human tissues, as a result of which they have a selective effect on the affected organs. This is due to the fact that the phosphonium group, due to its delocalized positive charge and high hydrophobicity, allows both known and new medicinal molecules conjugated to it in a certain way to easily penetrate not only through cell membranes, but also to overcome the high membrane potential of mitochondria (J. ZIELONKA et al., Mitochondria-targeted triphenylphosphonium-based compounds: syntheses, mechanism sofaction, and therapeutic and diagnostic applications, CHEMICAL REVIEWS, 2017, Vol. 117, No. 15, pp. 10043-10120). The distribution of charges provides more opportunities for the compound to orient itself within the target structure, thereby achieving the effectiveness of the drug. Therefore, the development of compounds and their forms with delocalized or separated charges has great potential.

Zwitterionic forms of the substance should be noted separately, since their advantages from the pharmacological point of view can be quite significant. Such compounds have a high dipole moment and dissolve very well in polar solvents, which can lead to increased bioavailability and acceleration of the onset of the therapeutic effect, as well as the possibility of introducing new routes of drug delivery, such as transbuccal, sublingual, rectal, inhalation or using transdermal therapeutic systems.

The technical results that the present invention is aimed at achieving are:

– increasing the solubility of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid;

– acceleration of the onset of the therapeutic effect of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid;

– maintaining the high efficiency of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid;

– increasing the long-term stability of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid (for 20 years);

– improving compressibility while maintaining good flowability without noticeable electrostatic phenomena of the pharmaceutical composition and dosage form containing the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid;

– effective antipyretic action;

– expansion of the arsenal of drugs for the treatment and/or prevention of rhinitis and pharyngitis;

– the possibility of obtaining a stable inhalation composition without adding carriers with non-respirable particle size, which are traditional for the state of the art;

– expanding the arsenal of antiviral agents;

– qualitative and/or quantitative changes in certain types of biological activity, including antioxidant, hypoglycemic, hypolipidemic, antihypoxic activities, as well as types of biological activity specified in RU 2141483 C1.

The technical results relate to both solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid and pharmaceutical compositions and medicinal products containing them.

New forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid

The stated problem is solved and the stated technical results are achieved thanks to new solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid. It was unexpectedly discovered that the said forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid contain a zwitterion of formula (II)

or its dimer of formula (III)

Zwitterion is a neutral molecule that contains spatially separated and opposite charges. In the zwitterion of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of formula (II), the carboxyl group is negatively charged (carboxylate ion), and the imidazole group is positively charged.

The zwitterionic structure was investigated by solid-state NMR 13C and X-ray diffraction methods and is fully confirmed by the materials of this application.

The term “new solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid” means forms containing 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid in the form of a zwitterion of formula (II) or a dimer of a zwitterion of formula (III).

The term solid form refers to crystalline and amorphous solid forms.

In one embodiment, the solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid comprises a dimer of formula (III) and belongs to a crystalline form characterized by a monoclinic crystal lattice syngony and the values of the edges of the crystal cell (in angstroms, Å): 8.71±0.50Å, 20.01±0.50Å and 12.16±0.50Å and an angle value (in degrees) of 93.10 ± 5.00 ^o . In a more preferred embodiment, the solid form has a layered structure, within each layer, the dimers of the zwitterions of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid are linked to each other by hydrogen bonds. Additionally, the crystal cell and the crystal lattice of the solid form can be characterized by the data of Figs. 1-3.

In another embodiment, the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic acid contains a zwitterion of formula (II) and belongs to the crystalline form characterized by monoclinic crystal lattice syngony and the values of the edges of the crystal cell (in angstroms, Å): 5.39±0.50, 10.08±0.50 and 19.85±0.50 and the value of the angle (in degrees) of 100.26 ± 5.00 ^o . In a more preferred embodiment, in the obtained form, the molecules of the zwitterions of 2-(imidazol-4-yl)ethanamide pentanedioic acid are linked to each other by hydrogen bonds in such a way that they are located approximately perpendicularly (if imaginary planes are drawn through the molecules), at an angle of approximately 90° (from 80 to 100 ^o ) relative to each other. Additionally, the crystal cell and crystal lattice of the solid form can be characterized by the data of Figs.4-6.

A crystal cell is a conventional parallelepiped built on the nodes of a crystal lattice, parallel translations of which in three dimensions allow the entire crystal lattice to be constructed. The parameters of a crystal lattice are the values of the edges ( a , b and c ) and the values of the angles between them (α, β and γ). In the monoclinic syngony of a crystal lattice, each crystal cell is characterized by three different edges ( a , b and c ), one angle different from a right angle, and two right angles.

The present invention also relates to solid forms comprising all possible mesomers of the zwitterion of formula (II) and the dimer of the zwitterion of formula (III), including:

Mesomers by imidazole group:
Mesomers by carboxyl group: Mesomers by amide group:

It will be clear to a person skilled in the art that all mesomers of the compound of formula (II) and formula (III), obtained according to the known rules of chemistry, also belong to the indicated structures.

A mesomer (mesomeric structure) is a structure in chemical resonance theory that arises from the conjugation of multiple bonds and/or unshared electron pairs in a molecule. In the structure of formula (II), the mesomeric effect is possible for the imidazole, carboxyl, and amide groups.

Taking into account the mesomeric effect, alternative designations of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid according to the present invention are:

Pentandioic acid imidazole ethanamide;

2-(Imidazol-5-yl)ethanamide pentanedioic-1,5 acid;

5-[2-(1H-Imidazol-4-yl)ethylamino]-5-oxopentanoic acid;

5-[2-(1H-Imidazol-5-yl)ethylamino]-5-oxopentanoic acid;

4-{[2-(1H-Imidazol-4-yl)ethyl]carbamoyl}butanoic acid;

4-{[2-(1H-Imidazol-5-yl)ethyl]carbamoyl}butanoic acid;

Method for obtaining new forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid

Glutaric anhydride and histamine or histamine salt can be used as starting materials for obtaining solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid. The method for obtaining solid forms includes the following stages:

Step 1 (optional step): Reaction of histamine dihydrochloride or another histamine salt with a base to form histamine in the form of a free amine;

Stage 2. Interaction of histamine with glutaric anhydride;

Stage 3. Treatment of the product obtained in stage 2 with mineral acid;

Step 4. Alkalinization of the obtained product (or addition of a base) until the precipitation of a solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid is complete, containing 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid as a zwitterion;

Step 5 (optional step): Purification of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid by known methods.

Steps 1-4 can be performed in one reactor (reaction flask) or in different reactors (reaction flasks) with intermediate isolation of products after each step.

A detailed description of each stage is provided below.

Stage 1. Interaction of histamine dihydrochloride with a base

The reaction of histamine dihydrochloride with a base is carried out to obtain histamine as a free base.

A solution of histamine dihydrochloride is prepared in any suitable solvent. A suitable solvent may be selected from an alcohol, ether, haloalkane, aromatic hydrocarbon or nitroalkane. Examples of alcohol are methyl alcohol (methanol), ethyl alcohol (ethanol) and isopropyl alcohol (isopropanol). Examples of ethers are diethyl ether (ethoxyethane), tetrahydrofuran (THF), dioxane. Examples of haloalkanes are chloroform, methylene chloride. Examples of aromatic hydrocarbons are benzene, toluene and xylenes. Examples of nitroalkanes are nitromethane and nitroethane. In a preferred embodiment, the amount of solvent is selected so as to ensure complete dissolution of histamine dihydrochloride. The concentration of the compound can be from 0.01 mol/L (mmol/mL) to 0.5 mol/L (mmol/mL), such as 0.01 mol/L, 0.05 mol/L, 0.1 mol/L, 0.2 mol/L, 0.25 mol/L, 0.3 mol/L, 0.4 mol/L, and 0.5 mol/L.

After dissolving the histamine dihydrochloride, a suitable base is added to the solution. Suitable bases include, for example, sodium hydroxide or potassium hydroxide. Strong amines, such as triethylamine, can also be used.

After completion of the neutralization reaction, the resulting amine can be isolated by known methods, for example, extraction followed by distillation of solvents. Histamine in the form of a free base can be introduced into the next stage without isolation. In addition, stage 1 can be carried out together with stage 2. For this, stage 2 is carried out in the presence of a suitable base, for example, in the presence of pyridine, triethylamine, morpholine.

If histamine free base is used as the starting compound, then step 1 is not necessary.

Stage 2. Interaction of histamine with glutaric anhydride

To carry out the reaction, a solution of histamine is prepared in any suitable solvent. A suitable solvent may be selected from an alcohol, ether, haloalkane, aromatic hydrocarbon or nitroalkane. Examples of alcohol are methyl alcohol (methanol), ethyl alcohol (ethanol) and isopropyl alcohol (isopropanol). Examples of ethers are diethyl ether (ethoxyethane), tetrahydrofuran (THF), dioxane. Examples of haloalkanes are chloroform, methylene chloride. Examples of aromatic hydrocarbons are benzene, toluene and xylenes. Examples of nitroalkanes are nitromethane and nitroethane. In a preferred embodiment, the amount of solvent is selected so as to ensure complete dissolution of histamine. The concentration of histamine can range from 0.01 mol/L (mmol/mL) to 0.5 mol/L (mmol/mL), such as 0.01 mol/L, 0.05 mol/L, 0.1 mol/L, 0.2 mol/L, 0.25 mol/L, 0.3 mol/L, 0.4 mol/L and 0.5 mol/L.

After the histamine has dissolved, glutaric anhydride is added to the solution. Glutaric anhydride is taken in an equimolar or excess amount. Glutaric anhydride can be added as a white solid without solvent or as a solution in a suitable solvent, such as the same solvent that was chosen to dissolve the histamine.

Preferably, the reaction is carried out at room temperature and with stirring. If the reaction is slow, the mixture can be heated to 30-50 ^o C or higher. The maximum temperature is determined by the boiling point of the solvent. If the reaction is too vigorous, the mixture can be cooled to 5-15 ^o C.

The reaction time is determined by the amounts of histamine and glutaric anhydride taken and can be from 0.1 hour to 24 hours, preferably from 1 to 6 hours, and even more preferably from 2 to 3 hours. The moment of completion of the reaction can be determined, for example, by thin-layer chromatography by the disappearance of the histamine spot. The moment of completion of the reaction can also be determined by the cessation of formation of the product sediment.

The reaction product can be isolated from the reaction mixture by known methods, such as filtration. The product can also be introduced into the next stage without intermediate isolation.

Stage 3. Treatment of sediment with mineral acid

The precipitate obtained in stage 2 is treated with an aqueous solution of mineral acid. Hydrochloric, sulfuric, phosphoric, hydrochloric, nitric and other strong acids can be selected as such an acid; in the preferred embodiment, the acid is hydrochloric acid.

The amount of acid is determined based on the amount of sediment being dissolved. For more complete dissolution, a small excess of acid can be taken (10-30% of the amount of sediment).

The permissible concentration of the acid is from 0.01 M (mol/L) to 5.00 M (mol/L), more preferably from 0.1 M to 0.2 M. The moment of completion of the reaction can be determined visually by the completion of the dissolution of the precipitate.

Stage 4. Alkalization of the resulting product

A suitable base is added to the solution obtained in step 3. In this case, the solution becomes cloudy and/or a precipitate forms in the form of crystals of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid containing a zwitterion of formula (II) or its dimer of formula (III). A suitable base can be an alkali (NaOH, KOH), soda (Na ₂ CO ₃ *10H ₂ O, Na ₂ CO ₃ , K ₂ CO ₃ , NaHCO ₃ ), amine (triethylamine, diethylamine, morpholine, pyridine, etc.).

To accelerate the precipitation of crystals, an auxiliary solvent, such as ethanol or methanol, can be added to the solution.

The reaction is carried out at room temperature or upon cooling to 5-15 ^o C. It is recommended to control the pH of the reaction mixture. The moment of completion of the reaction can be determined by the pH value of 5.0-6.5 of the reaction mixture, in the preferred embodiment the pH is 5.2-6.2. In another embodiment, the reaction is completed after the cessation of crystal precipitation.

The resulting crystals are isolated using known methods, for example, by filtration.

Stage 5. Product purification

The precipitate of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid can be further purified by known methods. For example, the precipitate can be washed with organic solvents (hexane, ether), after which it is further dried.

Pharmaceutical composition

A pharmaceutical composition is understood to mean a composition (mixture, composition, etc.) suitable for use in humans or animals, including an active pharmaceutical substance. The active pharmaceutical substance in the pharmaceutical composition includes an active substance - a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention or a mixture of solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention.

The mixture of solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention, in turn, contains a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid containing a zwitterion of formula (II)

and a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid containing a dimerzwitterion of formula (III)

The ratio of said solid forms can be selected in the range (by weight) from 1:10 to 10:1, preferably from 5:1 to 1:5, more preferably from 2:1 to 1:2, for example 1:1.

The active pharmaceutical substance can be used for the treatment and/or prevention of diseases caused by viral infections.

The present technical problem is solved and the said technical results are achieved thanks to a pharmaceutical composition for the treatment and/or prevention of diseases caused by viral infections, containing the above-defined active pharmaceutical substance, namely the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention or a mixture of solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention and at least one pharmaceutically acceptable excipient.

It will be understood by one skilled in the art that the pharmaceutical composition of the present invention will also include compositions containing one or more other active pharmaceutical substances, for example other antiviral pharmaceutical substances.

The terms “includes” and “contains” in the context of the present invention mean that the said pharmaceutical compositions (medicines, groups of components, etc.) include the following listed components/ingredients, but do not exclude the content of other components/ingredients.

In a preferred embodiment, the pharmaceutical composition is a composition in the form of a solid form. Examples of solid dosage forms are powders, granules, briquettes, capsules, tablets, dragees, etc. A person skilled in the art will understand that the pharmaceutical composition of the present invention can have any other form that ensures the preservation of the positive properties of the active pharmaceutical substance. In the most preferred embodiment, the solid form is a powder, granule, capsule or tablet. A powder is a solid non-dosed dosage form consisting of solid individual dry particles of varying dispersion, possessing the property of flowability. A capsule is a solid dosed or non-dosed dosage form, including a hard (usually gelatin) shell, inside which an encapsulate is enclosed, containing one or more active substances with or without the addition of excipients. A granule is a solid dosage form in the form of grains (aggregates of powder particles) of round, cylindrical or irregular shape, containing one or more active substances with the addition of excipients. A tablet is a solid dosage form, most often obtained by pressing powders or granules containing one or more active substances with or without the addition of excipients.

The quantitative content of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid in the pharmaceutical composition is selected from the range from 0.01 to 99.99 wt.%, in the preferred embodiment from 1.00 to 80.00 wt.%, more preferably from 10.00 to 60.00 wt.%, for example, 5.00 wt.%, 10.00 wt.%, 15.00 wt.%, 20.00 wt.%, 25.00 wt.%, 30.00 wt.%, 35.00 wt.%, 40.00 wt.%, 45.00 wt.%, 50.00 wt.%, 55.00 wt.%, 60.00 wt.%, from 10.00 wt.% to 20.00 wt.%; from 20.00 wt.% to 30.00 wt.%; from 30.00 wt.% to 40.00 wt.%; from 40.00 wt.% to 50.00 wt.%; from 50.00 wt.% to 60.00 wt.%, including all intermediate values in increments of 1.00 wt.%. For example, the value 23.00 wt.% is included as one of the values in the range from 20.00 wt.% to 30.00 wt.%.

In another preferred embodiment, the pharmaceutical composition comprises a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention in an effective amount.

The term "effective amount" in the context of the present invention refers to the amount of a pharmaceutical composition or drug that, when administered to a subject, is sufficient to effect such treatment on a disease, disorder or symptom. "Effective amount" may vary, for example, depending on the form in which the substance is present, the nature of the disease, disorder and/or symptoms of the disease or disorder, the severity of the disease, disorder and/or symptoms of the disease or disorder, the age of the subject to be treated, and/or the weight of the subject to be treated. The appropriate amount in each particular case will be obvious to a person skilled in the art or can be determined by routine experimentation.

In a preferred embodiment, the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid is contained in a pharmaceutical composition or a medicinal product and is used in a certain dosage. The dosage of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid according to the present invention can be in the range from 1 mg to 1000 mg per day, preferably from 20 mg to 100 mg, most preferably from 30 mg to 90 mg per day. The said dosage can be administered either as a single dose or divided into several doses during the day, for example, 2, 3 or 4 times per day.

The term "dosage" as used herein characterizes the content of one or more active substances in quantitative terms per unit dose, or unit volume, or unit mass in accordance with the dosage form, or for some types of dosage forms, the amount of active substance released from the dosage form per unit time.

The pharmaceutical composition of the present invention includes at least one pharmaceutically acceptable excipient, which is a carrier of active substances, providing the required volume/weight and the necessary characteristics of the drug in a certain dosage form. In a preferred embodiment, the pharmaceutical composition includes a pharmaceutically acceptable excipient, which is selected from the group consisting of a filler, a binder, a lubricant, a disintegrating agent, a sliding agent, a preservative, a flavoring agent and a coloring agent.

The term "filler" or "diluent" means auxiliary substances used to impart a given volume or weight to solid dosage forms. Starch, glucose, sucrose, lactose (milk sugar), basic magnesium carbonate, magnesium oxide, sodium chloride, sodium hydrogen carbonate, white clay (kaolin), gelatin, microcrystalline cellulose (MCC), methylcellulose (MC), sodium carboxymethylcellulose (Na CMC), calcium carbonate, dibasic calcium phosphate, glycine (aminoacetic acid), dextrin, amylopectin, ultra-amylopectin, sorbitol, mannitol, pectin, etc. can be used as fillers. The group is not limited to this list.

The term "binders" means substances included in the tablet mass to impart the required viscosity. Water, ethyl alcohol, starch paste, sugar syrup, solutions of carboxymethyl cellulose (CMC), oxyethyl cellulose (OEC), oxypropyl methyl cellulose (OPMC); polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), alginic acid, sodium alginate, gelatin, etc. are used as binding substances. The group is not limited to this list.

The term "lubricant" means auxiliary substances used in the technological process of tablet production at the pressing stage to improve the flowability of granules or powder by reducing friction between particles. The lubricant may be one or more of starch, talc, polyethylene oxide-4000, stearic acid, calcium and magnesium stearate, etc. The group is not limited to this list.

The term "lubricant" or "lubricant" means auxiliary substances that help reduce the friction force between the surface of the tablet and the walls of the punch tray in which the tablet is formed, used in the technological process of tablet production at the pressing stage. The lubricant may be one or more of magnesium stearate, calcium stearate, sodium stearyl fumarate, polyethyleneglycol (with a molecular weight of over 3350), sodium lauryl sulfate, talc, mineral oil, leucine and poloxamer, etc. The group is not limited to this list.

The term "disintegrating agent" means substances used to improve disintegration or dissolution by mechanically breaking up tablets in a liquid medium, which is necessary for the fastest release of the active substance. The disintegrating agent may be one or more of microcrystalline cellulose, sodium croscarmellose, crospovidone, sodium starch glycolate, starch, pectin, gelatin, amylopectin, ultra-amylopectin, agar-agar, alginic acid, potassium and sodium alginate, tween-80, etc. The group is not limited to this list.

Additional components, corrigents, are used to improve taste (sweetener) and smell (flavoring agent). These include, for example, sugar, cocoa, vanillin. Dyes (pigments) are used to improve the appearance of the pharmaceutical composition and dosage form. Examples of dyes are titanium dioxide, indigo carmine.

The amount, composition and form of the pharmaceutically acceptable excipient can be selected arbitrarily by a person skilled in the art, provided that the zwitterion of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of formula (II) or its dimer of formula (III) is completely or partially preserved.

In a preferred embodiment, the pharmaceutical composition according to the present invention comprises

a solid form of 2-(imidazol-4-yl)ethanamide of the present invention as an active substance and

mannitol (E421), sucralose - as auxiliary substances.

In an even more preferred embodiment, the pharmaceutical composition according to the present invention comprises

a solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid of the present invention in an amount of 0.09 g;

a mixture of mannitol (E421), sucralose, lemon lime flavoring – up to 3.0 g.

Routes of administration of the pharmaceutical composition of the present invention include, but are not limited to, oral, inhalation, topical, transdermal, sublingual and rectal routes. In a preferred embodiment, the pharmaceutical composition is administered orally or sublingually.

In some embodiments, the pharmaceutical composition of the present invention can be prepared using known conventional methods in the pharmaceutical field.

Medicine

The present technical problem is solved, and the said technical results are also achieved thanks to a medicinal product for the treatment and/or prevention of diseases caused by viral infections, containing a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention or a pharmaceutical composition of the present invention.

A medicinal product (drug) is a pharmaceutical composition in the form of a dosage form suitable for use in humans or animals.

In a preferred embodiment, the present invention relates to a drug in solid dosage form. Dosage forms of the present invention include, but are not limited to, tablets such as buccal tablets, effervescent tablets, film-coated tablets, sustained-release tablets, regular tablets, controlled-release tablets, orodispersible tablets, lyophilized fast-dissolving tablets, and the like; capsules such as hard capsules, soft capsules, gelatin capsules, and the like; injection powders such as lyophilized injection powders, regular injection powders, and the like; solutions, aerosols, sprays, transdermal drugs such as patches, granules, suppositories, and other dosage forms.

It will be clear to a person skilled in the art that the medicinal product of the present invention may have any form that ensures the preservation of the positive properties of the active pharmaceutical substance. In the most preferred embodiment, the solid form is a powder, granule, capsule or tablet.

In a preferred embodiment, the medicinal product according to the present invention comprises

a solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid of the present invention as an active substance and

mannitol (E421), sucralose - as auxiliary substances;

and is a powder.

In an even more preferred embodiment, the medicinal product according to the present invention comprises

a solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid of the present invention in an amount of 0.09 g;

a mixture of mannitol (E421), sucralose, lemon lime flavoring – up to 3.0 g of composition;

and is a powder.

The medicinal product of the present invention may be an active pharmaceutical substance and a pharmaceutical composition containing a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention. In a preferred embodiment, the medicinal product also includes at least one pharmaceutically acceptable excipient, however, its presence is not mandatory.

The quantitative content of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid in the medicinal product is selected from the range from 0.01 to 99.99 wt.%, in the preferred embodiment from 1.00 to 80.00 wt.%, more preferably from 10.00 to 60.00 wt.%, for example, 5.00 wt.%, 10.00 wt.%, 15.00 wt.%, 20.00 wt.%, 25.00 wt.%, 30.00 wt.%, 35.00 wt.%, 40.00 wt.%, 45.00 wt.%, 50.00 wt.%, 55.00 wt.%, 60.00 wt.%, from 10.00 wt.% to 20.00 wt.%; from 20.00 wt.% to 30.00 wt.%; from 30.00 wt.% to 40.00 wt.%; from 40.00 wt.% to 50.00 wt.%; from 50.00 wt.% to 60.00 wt.%. The stated content ranges mean that the solid form of the invention can contain each of the stated values within the stated range, including the extreme points. For example, a content of from 30 to 60 wt.% includes each value in the range from 30 to 60 wt.% in increments of 0.01 wt.%, for example, 43.21 wt.%.

The medicinal products of the present invention can be administered (without limitation) orally, parenterally, by inhalation, subcutaneously, intraperitoneally, locally, rectally. The therapeutic dosage of the medicinal product containing the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid, or the pharmaceutical composition of the present invention, or the pharmaceutical composition for the treatment of diseases caused by viral infections, which contains a therapeutically effective amount of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention, in patients can be adjusted depending on the therapeutic efficacy and bioavailability of the active ingredients in the body, the rate of their metabolism and elimination from the body, as well as depending on the age, sex and stage of the disease of the patient.

Inhalation composition

The present inventors have unexpectedly found that the solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention are suitable for the preparation of a medicinal product in the form of a dry powder for an inhaler (inhalation composition) without the addition of any auxiliary excipients.

The most common carrier for creating inhalation formulations is lactose, such as alpha-lactose monohydrate or beta-lactose anhydrous. Lactose as a carrier improves filling, dosing and delivery of the drug from the inhaler. To penetrate the lungs, the particle size of the active pharmaceutical substance should be in the range of 1-10 μm. To achieve such a small aerodynamic particle size, the active substance is processed by drying or micronization. However, micronized particles often form agglomerated powder that is difficult to dose. Mixing with lactose allows artificially breaking up agglomerates of the active substance and improving its use. It is assumed that at the outlet of the inhaler, the drug particles and lactose particles should separate (disaggregation). Large particles of lactose are non-respirable and settle in the oral cavity, while significantly smaller particles of the drug move into the respiratory tract under the influence of the air flow (V.V. Arkhipov and N.B. LAZAREVA, Principles of adequate inhalation, PRACTICAL PULMONOLOGY, 2018, 3, pp. 66-74). This approach was implemented in patent EA016363B1, which proposes an inhalation composition containing 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid and a biologically inert carrier using lactose as an example.

The distinctive advantages of the inhalation composition of the present invention are stability, minimization of electrostatic interactions and caking, preservation of particle size distribution during storage, and the absence of carriers with non-respirable particle size traditional in the prior art.

The inhalation composition of the present invention can be used not only for the treatment and/or prevention of diseases caused by viral infections, but also for the treatment and/or prevention of COPD, asthma and chronic bronchitis.

Use of a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid and a pharmaceutical composition and medicinal product containing it

Each solid form of 2-(imidazol-4-yl)ethanamide-1,5-pentanedioic acid of the present invention, a pharmaceutical composition containing it and a medicinal product of the present invention can be used to treat and/or prevent diseases caused by viral infections. In a preferred embodiment, viral infections include rhinovirus, respiratory syncytial virus, parainfluenza virus, influenza virus, including influenza A, B, C and D, adenovirus, noravirus, metapneumovirus, coronavirus, hantavirus, bocavirus, coxsackie virus, ECHO group viruses, enterovirus, rotavirus, herpes virus. In an even more preferred embodiment, viral infections are influenza type A virus. In another even more preferred embodiment, viral infections are SARS-CoV-2 coronavirus. Viral infections can be DNA and RNA viruses.

Each solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention, a pharmaceutical composition containing it and a medicinal product of the present invention can be used for the treatment and/or prevention of rhinitis, pharyngitis and sinusitis (rhinosinusitis).

Each solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention, a pharmaceutical composition containing it and a medicinal product of the present invention can be used as effective antipyretic agents. The antipyretic effect can be manifested in a decrease in body temperature during fever.

Each solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid and pharmaceutical composition containing it of the present invention can be used for the manufacture of a medicinal product of the present invention.

In one embodiment of the present invention, the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid of the present invention in the pharmaceutical composition or medicament of the present invention is administered in an amount of 1 mg to 1000 mg per day, preferably 20 mg to 100 mg, most preferably 30 mg to 90 mg per day. The said dosage can be administered either as a single dose or divided into several doses during the day, for example, 2, 3 or 4 times per day. The most preferred dosage options can be determined based on the age and/or body weight of the patient, as well as the nature of the symptoms exhibited.

Unless otherwise specified, all technical and specialized terms used in this context have their generally accepted meaning in the art.

Brief description of the drawings

Fig. 1 – Dimerzwitterion of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid in solid form;

Fig. 2 – Structure of the crystal cell of the solid form containing the dimerzwitterion of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid;

Fig. 3 – Packing of molecules in the crystal lattice of a solid form containing the dimerzwitterion of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid;

Fig. 4 – Zwitterion of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid in solid form;

Fig. 5 – Structure of the crystal cell of the solid form containing the zwitterion of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid;

Fig. 6 – Packing of molecules in the crystal lattice of a solid form containing the dimerzwitterion of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid;

Fig. 7 – General view of the spectrum of ¹³ C-CP/MAS of the solid form of the present invention;

Fig.8 – General view of ^{the 13} C - ¹ HHETCOR/MAS correlation;

Fig.9 – General view of the spectrum of ¹⁵ N-CP/MAS;

Fig.10 – Correlation of calculated and experimental chemical shifts ^{of 13} C and ¹⁵ N;

Fig.11 – Summary data on the distribution of particles in samples in the inhalation composition;

Fig.12 – Study of activity in the model of acute rhinosinusitis in rats;

* – differences are statistically significant compared to the intact group (p <0.05, 1-way Anova test);

& – differences are statistically significant compared to the control group (solvent) (p <0.05, 1-way Anova test);

Fig.13 – Study of activity in the model of non-infectious pharyngitis in rats;

* - differences are statistically significant compared to the intact group (p <0.05, 1-way Anova test);

& - differences are statistically significant compared to the control group (solvent) (p <0.05, 1-way Anova test);

Fig.14 – Study of activity in the model of febrile reaction in rats;

* - differences are statistically significant compared to the intact group (p <0.05, 1-way Anova test);

& - differences are statistically significant compared to the control group (solvent) (p <0.05, 1-way Anova test).

Implementation of the invention

The present invention is further illustrated, but not limited, by the following examples.

Example 1. Preparation of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid

A solution of 0.670 g glutaric anhydride in 10 ml THF was added dropwise with stirring to a solution of 0.500 g (4.5 mmol) of histamine in 10 ml of tetrahydrofuran (THF). The mixture was stirred at room temperature for 2 hours. The formed precipitate was filtered off and dissolved in a minimum amount of 0.1 M hydrochloric acid. To neutralize the resulting solution, a 5-fold amount of ethyl alcohol (by volume) was added to it, after which freshly distilled triethylamine was slowly added to the mixture until a pH of 5.2-5.7 was obtained. Upon reaching the desired pH value, the reaction product began to precipitate as colorless crystals. The addition of the base was stopped after the precipitation had ceased. The crystalline precipitate was filtered off and washed from traces of amine with ether cooled to 5 ^o C. The product was dried in air. Obtained: 0.640 g (63%) of a solid form containing dimerzwitterione 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid.

Example 2. Confirmation of zwitterionic structure by X-ray diffraction methods

X-ray diffraction analysis was performed on a BrukerQuest diffractometer. The experimental set of reflections was obtained from a single crystal at 100 K and Mo Kα radiation (λ = 0.71073). Using the Olex2 software (O.V.DOLOMANOV et al., OLEX2: a complete structuresolution, refinement and analysis program, JOURNALOFAPPLIEDCRYSTALLOGRAPHY, 2009, Vol.42, No.2, pp.339-341), the structure was solved using the SHELXL software package (G.M.Sheldrick, Crystal structure refinement with SHELXL, ACTA CRYSTALLOGRAPHICA SECTION C: STRUCTURAL CHEMISTRY, 2015, Vol.71, No.1, pp.3-8) and refined based on the least squares method. The figure of the decoded molecule is shown in Fig. 1.

From the obtained data it follows that a solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid was obtained, containing a dimerzwitterion of the following formula:

Crystallographic information for the unit cell _{of C20H30N6O6 (} M = _450.59 g/ _mol ): monoclinic syngony, space group _P21 /c, a = 8.7166 Å, b = 20.0076 Å, c = 12.1681 Å, α = 90°, β = 93.101 ^o , γ = 90°, V = 2118.99 ^Å3 , Z = 4, T = 140 K.

Additionally, the mutual arrangement of molecules in the crystal cell and crystal lattice was analyzed. The results are shown in Fig. 2 and Fig. 3, respectively. It was found that the obtained form has a layered structure, within each layer, the dimers of zwitterions of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid are linked by hydrogen bonds. There are no hydrogen bonds of dimers between adjacent layers, and crystallographic planes separating individual layers can be clearly distinguished (see Fig. 3).

Example 3. Preparation of a solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid

A solution of 0.670 g glutaric anhydride in 10 ml dioxane was added dropwise with stirring to a solution of 0.500 g (4.5 mmol) of histamine in 10 ml of dioxane. The mixture was stirred at room temperature for 2 hours. The formed precipitate was filtered off and dissolved in a minimum amount of 0.1 M hydrochloric acid. To neutralize the resulting solution, a 5-fold amount of ethyl alcohol (by volume) was added to it, after which a 10% sodium carbonate solution was slowly added to the mixture until a pH of 6.0-6.5 was obtained. Upon reaching the desired pH value, the reaction product began to precipitate as colorless crystals. The addition of the base was stopped after the precipitation had ceased. The crystalline precipitate was filtered off and washed from traces of amine with hexane cooled to 5 ^o C. The product was dried in air. Obtained: 0.812 g (80%) of zwitterion 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid

Example 4. Confirmation of the zwitterionic structure by X-ray diffraction and solid-state NMR methods

X-ray diffraction analysis was performed on a BrukerQuest diffractometer. The experimental set of reflections was obtained from a single crystal at 100 K and Mo Kα radiation (λ = 0.71073). Using the Olex2 software (O.V.DOLOMANOV et al., OLEX2: a complete structure solution, refinement and analysis program, JOURNAL OF APPLIED CRYSTALLOGRAPHY, 2009, Vol. 42, No. 2, pp. 339-341), the structure was solved using the SHELXL software package (G.M.SHELDRICK, Crystal structure refinement with SHELXL, ACTA CRYSTALLOGRAPHICA SECTION C: STRUCTURAL CHEMISTRY, 2015, Vol. 71, No. 1, pp. 3-8) and refined based on the least squares method. The figure of the decoded molecule is shown in Fig. 4.

From the obtained data it follows that a solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid was obtained, containing a zwitterion of the following formula:

Crystallographic information for C ₁₀ H ₁₅ N ₃ O ₃ ( M = 225.24 g/mol): monoclinic syngony, space group P2 ₁ /c, a = 5.3967 Å, b = 10.0809 Å, c = 19.8540 Å, α = 90 ^o , β = 100.258 ^o , γ = 90 ^o , V = 1062.88 Å ³ , Z = 4, T = 140 K.

Additionally, the mutual arrangement of molecules in the crystal cell and the crystal lattice was analyzed. The results are shown in Fig. 5 and Fig. 6, respectively. In a separate cell, zwitterion dimers were not visible. In addition, it was found that in the obtained form, the molecules of zwitterions of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid are linked to each other by hydrogen bonds in such a way that they are located at an angle of approximately 90° (80-100°) relative to each other (see Fig. 6).

The solid-state NMR spectra were recorded using the cross-polarization/magic angle spinning (CP/MAS) technique. Solid-state NMR experiments were recorded on a Bruker 400 MHz spectrometer. Samples were spun at 11 to 14 kHz under magic angle spinning (MAS) using ZrO ₂ rotors. All 13C and 15N spectra were recorded under proton decoupling conditions.

In solid-state NMR 1H all signals were strongly broadened due to the experimental features. Therefore, to facilitate the assignment of hydrogen atom nuclei signals, the HETCOR method was used, which consists of obtaining a correlation of ¹ H and ¹³ C chemical shifts.

In solid-state ^13C NMR, intense peaks were observed at 179, 173, 135, 129, 117, 36, 33, 24, and 21 ppm. In addition, relatively weak peaks were present at 258, 254, 252, 214, 208, 197, 98, 95, 93, 55, and 49 ppm, which represented rotation bands of the main isotropic peaks. The spectrum is shown in Fig. 7. The spectrum recorded by the HETCOR method is shown in Fig. 8. Similar signals were also observed for the solid form according to Example 2.

In solid-state NMR ^15N intense peaks were observed at 123, 175 and 193 ppm, and the spectrum itself is shown in Fig. 9. The calculated chemical shifts of the ^13C and ^15N nuclei are presented in Table 1. The general appearance of the molecule and the numbering of the atoms are shown in Fig. 4.

Next, the chemical shifts of the maxima were calculated using quantum chemistry methods. The structure in Fig. 4, refined using powder diffraction data, was used as a starting point for the quantum chemical calculation. To improve the convergence of the calculation and experimental data, the atomic coordinates were pre-optimized.

For direct calculation of chemical shifts of nitrogen and carbon atoms the program VASP 5.4.1 was used, the GI-PAW method (see G.KRESSE and J.HAFNER, Ab initio molecular dynamics for liquid metals, PHYSICAL REVIEW B, 1993, Vol.47, No.1, p.558; G.KRESSE and J.HAFNER, Ab initio molecular-dynamics simulation of the liquid-metal–amorphous-semiconductor transition in germanium, Physical Review B, 1994, Vol.49, No.20, pp.14251-14269; G.KRESSE and J.FURTHMÜLLER, Efficient iterative schemes for ab initio total-energy calculations using a plane-wave basis set, PHYSICAL REVIEW B, 1996, Vol.54, No.16, p.11169; G.KRESSE and J.FURTHMÜLLER, Efficiency of ab-initio total energy calculations for metals and semiconductors using a plane-wave basis set, COMPUTATIONAL MATERIALS SCIENCE, 1996, Vol.6, No.1, pp.15-50; G.KRESSE and D.JOUBERT, From ultrasoft pseudopotentials to the projector augmented-wave method, PHYSICAL REVIEW B, 1999, Vol.59, No.3, p.1758). In this case, a grid with 16 k-points in the independent part of the Brillouin zone was used to solve the Kohn-Sham equations. The convergence threshold in the self-consistency procedure for determining the energy was set as 10 ^-10 eV. The orthorhombic modification of SiMe ₄ (see AKWOLF et al. Predicted crystal structures of tetramethylsilane and tetramethylgermane and an experimental low-temperature structure of tetramethylsilane, ACTA CRYSTALLOGRAPHICA SECTION B: STRUCTURAL SCIENCE, 2010, Vol. 66, No. 2, pp. 229-236) and the crystal structure of ammonia (see R. BOESE et al. Single-crystal structure and electron density distribution of ammonia at 160 k on the basis of X-ray diffraction data, THE JOURNAL OF PHYSICAL CHEMISTRY B, 1997, Vol. 101, No. 30, pp. 5794-5799) were used as standards. For both standards, quantum chemical calculations with geometry optimization were also performed and the magnetic shielding tensors of atomic nuclei were calculated.

Table 1. Calculated and experimental solid-state chemical shifts

Atom Calculation, m.d. Experiment, m.d. N4 128.8 122.9 N5 182.9 175.2 N6 193.9 193.0 C7 122.1 120.3 C8 138.0 132.3 C9 35.5 39.1 C10 22.6 24.0 C11 39.0 36.1 C12 27.4 26.9 C13 176.6 175.8 C14 38.7 39.1 C15 135.5 138.3 C16 185.3 181.7

The most significant differences between the calculation and the experiment are observed for the N4 and N5 atoms. Direct comparison of the chemical shift values allows us to conclude that the quantum-chemical calculation somewhat overestimates the strength of the NH…O hydrogen bonds. Apparently, strengthening of these bonds can lead to some local transfer of electron density from the NH group to the carbonyl oxygen atom and a shift to a strong field of all ¹⁵ N signals.

The agreement between the calculation and experiments in the case of ¹³ C chemical shifts is noticeably better. The greatest discrepancy is noted for the C8 atom located between the nitrogen atoms N5 and N6. The nature of this deviation is apparently the same as in the case of ¹⁵ N chemical shifts and is due to the strengthening of hydrogen bonds according to the results of the quantum chemical calculation compared to the experiment. The remaining ¹³ C chemical shifts are reproduced in the quantum chemical calculation within 3.0-3.5 ppm.

The comparison of the calculated and experimental data is presented in Fig. 10 and demonstrates the presence of a linear correlation. The calculated values of R ² and standard error are 0.998 and 1.69 ppm. The confidence interval is 5.76 ppm.

Measurement of ¹³ C and ¹⁵ N chemical shifts by NMR and quantum chemical calculation of these parameters showed that the composition of the solid form sample in the form of a zwitterion dimer is completely consistent with the structure established using the powder X-ray diffraction method.

Example 5. Obtaining a pharmaceutical composition and a dosage form in the form of capsules

To prepare the pharmaceutical composition and dosage form, the following were used:

Composition 1:

Solid form of 2-(imidazol-4-yl)ethanamide pentanedio- voy-1.5 acid obtained according to example 1 60.00 mg lactose monohydrate 60.00 mg potato starch 23.72 mg colloidal silicon dioxide 1.47 mg magnesium stearate 1.47 mg

Composition 2:

Solid form of 2-(imidazol-4-yl)ethanamide pentanedio- voy-1.5 acid obtained according to example 3 60.00 mg lactose monohydrate 60.00 mg potato starch 23.72 mg colloidal silicon dioxide 1.47 mg magnesium stearate 1.47 mg

The dosage form in the form of hard capsules was obtained by weighing and sifting the substances, mixing them and encapsulating them using a machine.

Example 6. Obtaining a pharmaceutical composition and dosage form in powder form

To prepare the pharmaceutical composition and dosage form, the following were used:

Composition 3:

Solid form of 2-(imidazol-4-yl)ethanamide pentanedio- voy-1.5 acid obtained according to example 1 90.00 mg a mixture of mannitol (E421), sucralose, lemon lime flavor up to 3.0 g.

Composition 4:

Solid form of 2-(imidazol-4-yl)ethanamide pentanedio- voy-1.5 acid obtained according to example 3 90.00 mg a mixture of mannitol (E421), sucralose, lemon lime flavor up to 3.0 g

The dosage form in powder form was obtained by weighing and mixing the above ingredients.

Example 7. Obtaining an active pharmaceutical substance in powder form

To 100 mg of the solid form according to example 1, 10 mg of the solid form according to example 3 were added. The resulting mixture was thoroughly mixed, weighed and placed in a weighing bottle with a tightly closing lid. 110 mg of the active pharmaceutical substance of the present invention were obtained.

To 100 mg of the solid form according to example 3, 10 mg of the solid form according to example 1 were added. The resulting mixture was thoroughly mixed, weighed and placed in a weighing bottle with a tightly closing lid. 110 mg of the active pharmaceutical substance of the present invention were obtained.

To 50 mg of the solid form according to example 1, 50 mg of the solid form according to example 3 were added. The resulting mixture was thoroughly mixed, weighed and placed in a weighing bottle with a tightly closing lid. 100 mg of the active pharmaceutical substance of the present invention were obtained.

Example 8. Obtaining an inhalation dosage form

Weighed portions of 200 mg of the solid form according to example 1 (sample A), 200 mg of the solid form according to example 3 (sample B) and the form obtained according to prototype RU 2141483 (sample C) were homogenized to a respirable size of 2-3 µm.

The type of grinding using a ball mill was selected. This type of mill uses the principle of high-energy impacts of grinding balls.

Among the equipment that uses this type of grinding, planetary ball mills are the simplest and most reliable, while at the same time they provide efficient grinding.

The grinding cups with the material to be ground and the balls are additionally rotated on a rapidly rotating planetary disk in the opposite direction around their own axis. In this position, the centrifugal force at a certain speed causes the material to be ground and the balls to separate from the inner wall of the grinding cup. In this case, the grinding balls cross the cup at maximum speed and grind the material to be ground on the opposite wall of the cup both by crushing and as a result of the balls colliding with each other.

A Pulveresetta 6 planetary ball mill, Fritsch, Germany, was used for grinding. With 500 ml grinding cups. The balls were 5 mm in diameter and made of zirconium oxide (ZrO ₂ ). Test measurements were performed on sample A.

Several experiments were conducted to select the optimal grinding parameters. The effect of time on the degree of grinding was investigated. To achieve the specified particle size, it was decided to increase the grinding time.

Table 2. Parameters of the sample grinding experiment.

Experience No. Amount of API, g Number of balls, pcs. Speed, rpm Time, min 1 150 2000 150 15 2 200 2000 150 15 3 250 2000 150 15 4 200 2000 175 15 5 200 2000 200 15 6 200 2000 175 15 7 200 2000 175 20 8 200 2000 175 30

Summary data on particle distribution in the samples are shown in Fig. 11. In experiment 8, a particle size of less than 3 μm was obtained for sample A. 10% of the particles were less than 0.7 μm (D ₁₀ ), 50% of the particles were less than 2.1 μm (D ₅₀ ) and 90% were less than 2.9 μm (D ₉₀ ). The particle sizes were determined on a MASTERSIZER 3000 laser diffraction particle analyzer. Samples B and C were ground under similar conditions.

The obtained samples were placed in bottles with tightly closing lids and stored at room temperature.

After two weeks, the obtained samples were re-examined using a MASTERSIZER 3000 laser diffraction particle analyzer. Samples A and B retained their respirable particle size distributions during storage.

Sample C contained particles larger than 100 µm. Without auxiliary excipients, this form is not suitable for use in an inhaler.

Example 9. Study of the anti-inflammatory activity of solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid in a model of carrageenan paw edema in rats.

A series of experiments were conducted on rats, which were randomly divided into groups, with 5 animals in each group. The data in the control were pooled. A model of carrageenin-induced paw edema in Wistar rats was used for the studies. The edema was induced by subplantar administration of 0.1 ml of a 1% aqueous solution of λ-carrageenin into the hind limb of animals in the experimental and control groups. Animals in the experimental groups received the test compounds orally using a probe immediately after the induction of carrageenin edema. The solid form of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid of the present invention, obtained according to Example 1, was administered to the animals at a dose of 10 mg/kg. Similarly, animals in the control group received the appropriate amount of placebo. Edema was assessed by measuring the foot volume using a mechanical oncometer before the administration of flagogen (λ-carrageenin), two hours after its administration, and four hours after its administration. The degree of edema was judged by the difference in volume between the edematous foot and the foot before inflammation induction. Anti-inflammatory activity was determined using the formula A = (ΔV _k -ΔV _e /ΔV _k ) · 100%, where A is the anti-inflammatory activity in percent; ΔV _e and ΔV _k are the difference between the edematous and normal feet in animals of the experimental and control groups.

The degree of reduction in foot swelling (average value for the group) after 2 hours of the study was 40% higher in the experimental group than in the control group.

Similar results were achieved for the solid form obtained in Example 3.

The presented data indicate that solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid according to the present invention exhibit anti-inflammatory activity.

Example 10. Study of storage stability of new solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid

The stability of solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid was assessed by the accelerated aging method according to OFS.1.1.0009.15 Shelf life of medicinal products, State Pharmacopoeia of the Russian Federation. At elevated temperatures, as a rule, the physicochemical processes occurring in medicinal products are accelerated, leading over time to undesirable changes in quality. Thus, at elevated temperatures, the period of time during which the controlled quality indicators of the medicinal product remain within acceptable limits (experimental shelf life) is artificially reduced.

The proposed storage conditions for solid forms of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid were chosen to be 10 years at a temperature not exceeding 25°C.

The shelf life ( S ) at the storage temperature ( t _хр ) is related to the experimental shelf life ( S _E ) at the elevated experimental storage temperature ( t _e ) by the Van't Hoff equation:

,

where A is the temperature coefficient, which is taken to be equal to 2.5.

The storage conditions selected for the accelerated aging experiment were 40 days at 75°C. To conduct the study, a sample of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid was left in a climatic chamber for 40 days at 75°C. The composition of the resulting sample was analyzed using high-performance liquid chromatography. The amount of impurities in the resulting chromatogram did not exceed 0.5% for each form, which confirms their stability.

Example 11. Activity study in the rat model of acute rhinosinusitis

Experimental methodology .The experimental group included animals (Wistar male rats) with no signs of abnormal appearance, so that the individual weight value deviated from the average value within the sex by no more than ±10%. Acute rhinosinusitis was induced by a single intranasal administration of 20 μl of 7.5% formalin into each nasal passage. The solid form according to Example 1 (hereinafter referred to as the active pharmaceutical substance or APS) in doses of 1.8, 18 mg/kg was administered intragastrically, daily, once a day, starting 24 hours after the formalin administration, the last administration occurred on the 7th day.

On the 8th day, the animals were anesthetized by intraperitoneal administration of a mixture of Zoletil and Xylazine (1:5) in a volume of 1 ml/kg. Cannulas were inserted through the trachea into the nasopharynx and washed with a syringe with a physiological solution in a volume of 2 ml at such a speed that the solution flowed out of the nose drop by drop. Using a Goryaev chamber, the absolute number of cells in the lavage was determined (in three repetitions). Nasal lavage was centrifuged for 10 minutes and smears were prepared from the cell sediment, which were fixed in methanol and stained according to Romanovsky-Giemsa. The relative number of neutrophils and macrophages was determined from the smears under a microscope. The count was made per 300 cells.

Descriptive statistics were used for all data: the arithmetic mean (Mean) and standard error of the mean (SEM) were calculated. The Shapiro-Wilk test was used to check the normality of the distribution of the values obtained during the experiment. In case of a non-normal distribution, 1-way ANOVA (with Dunn's post-analysis) was used to evaluate intergroup differences. In case of a normal distribution, 1-way ANOVA (with Tukey's post-analysis) was used to compare several groups. Statistical analysis was performed using the Graph Pad Prism. 9.0 program. Differences were determined at p<0.05.

The experimental conditions are presented in Table 2, and the results of the study are shown in Fig. 12.

Table 3. Activity study in the model of acute rhinosinusitis in rats

Group Model Administration of drugs Euthanasia Number of animals Intact 8th day of the study 10 Control (solvent) 20 µl of 7.5% formalin in each nasal passage, single intranasal Intragastrically, daily, once a day, starting 24 hours after the administration of formalin, the last administration is
on the 7th day 16 AFS (1.8 mg/kg) 10 AFS (18 mg/kg) 10

The study conducted shows that the solid form according to example 1 can be used for the treatment or prevention of acute rhinosinusitis.

Example 12. Activity study in a non-infectious pharyngitis model in rats

The non-infectious pharyngitis model in rats was implemented according to the standard technique (see G.L.VISWANATHA et al. Novel experimental model of non-infectious pharyngitis in rats, JOURNAL OF PHARMACOLOGICAL AND TOXICOLOGICAL METHODS, 2014, Vol. 69, No. 2, pp. 189-195). Animals (male Wistar rats) with no signs of abnormalities in appearance were selected for the experimental group in such a way that the individual weight value deviated from the average value within the sex by no more than 10%. Rats were anesthetized with sodium thiopentone (50 mg/kg, intraperitoneally) and a cannula with a RenaSil® silicone rubber tube (SIL 037, Braintree Scientific, Inc., Braintree, MA) was inserted into the external jugular vein. Evans blue dye (EB) was administered to all animals at a dose of 30 mg/kg intravenously through a catheter; 10 min after the administration of EB dye, formalin (formaldehyde solution at a concentration of 30%) was applied to the surface of the pharyngeal mucosa as follows: the tongue was slightly pulled out, the pharyngeal area was opened deeply using blunt forceps, and formalin (50 μl) was carefully applied with a sterile cotton swab three times for 5 sec with each application. In the intact group, physiological saline was used. The solid form according to Example 3 (hereinafter referred to as the active pharmaceutical substance or APS) at doses of 1.8, 9, 18 mg/kg was administered intragastrically 24 and 1 h before the application of formaldehyde.

Animals were sacrificed by exsanguination 60 minutes after formaldehyde application. The head of each rat was perfused with heparinized saline (40 U/ml) to remove intravascular EG dye.

The efficacy in treating pharyngitis was assessed using the Evans blue (EB) exudation test. EB dye was extracted from tissue into formamide at 55°C for 24 hours, and the absorbance was determined spectrophotometrically at 620 nm. The amount of EB dye in the tissue was calculated using a standard curve for EB dye and expressed as micrograms of dye per milligram of wet tissue weight (μg/mg).

Descriptive statistics were used for all data: the arithmetic mean (Mean) and standard error of the mean (SEM) were calculated. The Mann-Whitney test was used to compare groups. Statistical analysis was performed using the Graph Pad Prism. 9.0 program. Differences were determined at p<0.05.

The experimental conditions are presented in Table 3, and the results of the study are shown in Fig. 13.

Table 4. Activity study in the non-infectious pharyngitis model in rats

Group Administration of drugs Euthanasia Number of animals Intact (solvent)   1 hour after formalin application 10 Control (solvent) 24 and 1 hour before
application of formalin 10 AFS (1.8 mg/kg) 10 AFS (9 mg/kg) 10 AFS (18 mg/kg) 10

The study conducted shows that the solid form according to example 3 can be used for the treatment or prevention of non-infectious pharyngitis.

Example 13. Study of activity in a model of febrile reaction in rats

The model of febrile reaction in rats was implemented according to the standard method (F. ZHANG et al., Metabolomic study of the fever model induced by baker's yeast and the antipyretic effects of aspirin in rats using nuclear magnetic resonance and gas chromatography–mass spectrometry, JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS, 2013, Vol. 81, pp. 168-177). Animals (male Wistar rats) with no signs of abnormalities in appearance were selected for the experimental group, so that the individual weight value deviated from the average value within the sex by no more than ±10%. Wistar rats were subcutaneously injected with a 20% suspension of baker's yeast (12 ml/kg). The solid form according to Example 1 (hereinafter referred to as the active pharmaceutical substance or APS) at a dose of 1.8 mg/kg was administered intragastrically, once 14 hours after the introduction of the yeast. Rectal temperature was measured with an electric thermometer (AT-12R PHYSITEMP, made in Israel) before the introduction of the pyrogen and 14, 16, 17, 18, 19, 20, 24, 36, 48, 60 and 72 hours after it. The results were expressed as an increase in temperature relative to the values before the introduction of the pyrogen.

Descriptive statistics were used for all data: the arithmetic mean (Mean) and standard error of the mean (SEM) were calculated. The Shapiro-Wilk test was used to check the normality of the distribution of the values obtained during the experiment. In case of a non-normal distribution, 1-way ANOVA (with Dunn's post-analysis) was used to evaluate intergroup differences. In case of a normal distribution, 1-way ANOVA (with Tukey's post-analysis) was used to compare several groups. Statistical analysis was performed using the Graph Pad Prism. 9.0 program. Differences were determined at p<0.05.

The experimental conditions are presented in Table 4, and the results of the study are shown in Fig. 14.

Table 5. Study of activity in the febrile reaction model in rats

Group Administration of drugs Number of animals Intact   50 Control (solvent) Intragastric,
14 hours later
yeast introduction 50 AFS (1.8 mg/kg) 50

Claims (16)

1. A solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid containing a zwitterion dimer of formula (III) characterized in that the solid form refers to a crystalline form characterized by a monoclinic crystal lattice syngony and the values of the edges of the crystal cell (in angstroms, ): 8.71±0.50, 20.01±0.50 and 12.16±0.50 and the angle value (in degrees) 93.10±5.00°. 2. A solid form according to claim 1, characterized in that the solid form has a layered structure, within each layer the dimers of zwitterions of 2-(imidazol-4-yl)ethanamide of pentanedioic-1,5 acid are linked to each other by hydrogen bonds. 3. A method for obtaining a solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid according to any one of claims 1 or 2, comprising reacting histamine with glutaric anhydride and treating the resulting product with a mineral acid at a pH of 5.2-5.7, followed by alkalization or addition of a base until the precipitation of the solid form of 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid is complete, containing 2-(imidazol-4-yl)ethanamide pentanedioic-1,5 acid in the form of a dimer of formula (III). 4. The method according to claim 3, further comprising the step of obtaining histamine by reacting histamine dihydrochloride with a suitable base. 5. The method according to claim 3, further comprising adding a solvent selected from methanol or ethanol at the alkalization stage. 6. A pharmaceutical composition for the treatment and/or prevention of diseases caused by viral infections, containing a solid form according to any one of claims 1 or 2, and at least one pharmaceutically acceptable excipient, wherein the viral infections are parainfluenza virus, influenza virus, including influenza A, B, C and D. 7. A pharmaceutical composition according to claim 6, containing a solid form of 2-(imidazol-4-yl)ethanamide according to any one of claims 1 or 2 as an active substance and mannitol (E421), sucralose as excipients. 8. A medicinal product in solid form for the treatment and/or prevention of diseases caused by viral infections, containing a solid form according to any one of claims 1 or 2 or a pharmaceutical composition according to any one of claims 6 or 7, wherein the viral infections are parainfluenza virus, influenza virus, including influenza A, B, C and D. 9. The medicinal product according to item 8, characterized in that the solid form is a powder, granule, capsule or tablet. 10. The medicinal product according to claim 8 in the form of a powder, containing a solid form of 2-(imidazol-4-yl)ethanamide according to any one of claims 1 or 2 as an active substance and mannitol (E421), sucralose as excipients. 11. A medicinal product according to claim 8 in the form of a powder for inhalation, containing a solid form containing a solid form of 2-(imidazol-4-yl)ethanamide according to any one of claims 1 or 2, characterized in that said medicinal product does not contain auxiliary substances. 12. The medicinal product according to claim 8 in the form of capsules for inhalation, containing a solid form of 2-(imidazol-4-yl)ethanamide according to any one of claims 1 or 2, characterized in that said medicinal product does not contain auxiliary substances. 13. The medicinal product according to claim 8, characterized in that it includes at least one pharmaceutically acceptable excipient. 14. Use of a solid form according to any one of paragraphs 1 or 2 for the treatment or prevention of diseases caused by viral infections, wherein the viral infections are parainfluenza virus, influenza virus, including influenza A, B, C and D.