RU2815401C2 - Bisamide derivative of dicarboxylic acids, its use, pharmaceutical composition based on it, methods for its obtaining - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: pharmaceutical composition for the prevention and/or treatment of inflammatory diseases, comprising an effective amount of a compound of formula 11 or a pharmaceutically acceptable salt thereof. The invention also relates to a medicinal product for the prevention and/or treatment of inflammatory diseases, including a compound of formula 11.

11

EFFECT: compound with the ability to complex or chelate metal ions that may find use as a means for the prevention and/or treatment of inflammatory diseases.

2 cl, 5 tbl, 9 ex

Description

The invention relates to new biologically active compounds, derivatives of dicarboxylic acid bisamides or their pharmaceutically acceptable salts, with the ability to complex or chelate metal ions, as well as their use as a means of antioxidant action, a means for the prevention and/or treatment of cardiovascular, viral, oncological, neurodegenerative, inflammatory diseases, diabetes, gerontological diseases, diseases caused by microorganism toxins, as well as alcoholism, alcoholic cirrhosis of the liver, anemia, porphyria tarda, poisoning with transition metal salts.

State of the art

Metal ions play an important role both in the normal functioning of cells and the whole organism, and in the development of pathologies.

There are known drugs that exert their effect due to the ability to chelate metal ions. In this regard, a search is currently underway for non-toxic compounds that can highly efficiently and selectively chelate metal ions and are suitable for biomedical use.

Compounds with chelating properties were found among various classes of compounds: mono- and dithiols, disulfides, azo compounds, nitrosoaromatic compounds, polyaminocarboxylic acid derivatives, thiosemicarbazone, pyridoxal isonicotinoylhydrazone, quinoline, adamantane, pyrogallol, phenanthroline, thiopyrophosphates and others. In addition, they are noted among other natural compounds, such as, for example, carnosine, phytin, pectin. Of greatest interest are compounds that have several functional groups that can act as electron donors during complex formation. In this regard, such compounds may be ligands that specifically interact with ions of a metal or group of metals.

Currently widely known complexing agents are derivatives of polyaminocarboxylic acids (for example, EDTA), D-penicillamine, and polycyclic cryptands, which are successfully used for heavy metal poisoning. Deferroxamine is used as an iron chelator in some iron overload conditions and hematochromatosis. In addition, it is possible to use chelators for pathologies associated with excess Ca, for example, arthrosis, atherosclerosis, and kidney stones. Chelation therapy is also known to prevent the deposition of cholesterol and restore its level in the blood, lower blood pressure, avoid angioplasty, suppress unwanted side effects of certain heart medications, remove calcium from cholesterol plaques, dissolve blood clots and restore the elasticity of blood vessels, normalize arrhythmia, and prevent aging. , restores the strength of the heart muscle and improves heart function, increases intracellular potassium content, regulates mineral metabolism, is useful in the treatment of Alzheimer's disease, prevents cancer, improves memory and exhibits many other positive effects. However, strong chelators currently used in chelation therapy usually have toxic effects, which manifest themselves mainly in damage to the mucous membrane of the small intestine and impaired renal function. In some cases, rapid administration of large quantities of known chelators may impair muscle excitability and blood clotting. In addition, strong chelators can interact with beneficial bioelements (Na, K, Ca, Mg, Ca), and can also change the activity of vital metalloenzymes [Zelenin K.N. "Complexons in medicine", Soros Educational Journal, 2001, vol. 7, no. 1, pp. 45-50].

In this regard, an active search is being carried out for new highly effective chelators with a chelating ability sufficient to produce a biological effect in vivo , without side effects.

The idea of using chelation therapy in the case of viral diseases such as HIV, human papilloma, herpes, hepatitis C and others is especially relevant.

A promising target for this seems to be the zinc-binding sites in the structure of viral proteins - the so-called “zinc fingers”.

Several compounds have now been identified that affect the zinc fingers of important proteins of these viruses.

The article by Andreas J.K., Boorganic & Medicinal chemistry, 2003, v.11, p.4599-4613, describes an adamantane derivative with the trivial name bananain, which is a chelator of zinc ions. It is thought to be chemically suited to remove zinc from the HIV NCp7 protein. An aza derivative with the above-mentioned mechanism of action, azadicarbonamide, is in stage I/II clinical trials against progressive AIDS.

Rice WG, Schaeffer CA, Harten B., Nature, 1993, v.4, p.473-475, disclosed 3-nitrosobenzamide, which removes zinc from the NCp7 protein of HIV, inhibiting HIV replication and its pathogenicity in vitro and in vivo .

The zinc finger region of the E6 protein of the human papillomavirus was also selected as a drug target. This virus is a possible mediator in the etiology of cervical carcinoma.

The article Beerheide W., Bernard H.-U., Tan Y.-J., Ganesan AJ, National Cancer Institute, 1999, v.91, No. 14, 1211-1220, describes in vitro tests of aza compounds, disulfide and nitrosoaromatic derivatives . Compounds such as 4,4'-dithiodimorpholine have been shown to provoke the release of zinc ions. As a result, changes in the structure of the viral protein and disruption of its functions associated with the biology and pathology of the human papillomavirus were observed. However, clinical trials of these compounds in this regard have not yet been completed, and their effectiveness can only be judged on the basis of in vitro studies.

The above compounds are considered promising for the development of drugs against cervical cancer, genital warts and latent human papillomavirus infections of the genital organs. Hepatitis C virus is one of the most widespread human pathogens. Modern therapy for hepatitis C is based almost exclusively on the use of interferon, as well as its combination with a nucleoside analogue - ribavirin [Kozlov M.V., Polyakov K.M., Ivanov A.V., Biochemistry, 2006, v. 71, No. 9, pp. 1253-12594]. It should be noted that this therapy is not very effective.

Regarding the development of therapeutic agents against the hepatitis C virus, one of the targets is the NS3-serine proteinase, in maintaining the stability of the structure of which the zinc region plays an important role [Andrea Urbani, Renzo Bazzo, Maria Chiara Nardi, Daniel Oscar Cicero, Raffaele De Francesco, J Biol. Chem, 1998, v.273, No. 30, p. 18760-18769]. Inhibition or modification of its activity by the use of zinc-extracting compounds has been assessed in some literature as a promising strategy for the management of hepatitis C virus disease.

In the article Timothy L. Tellinghuisen, Matthew S. Paulson, Charles M. Rice, J. Virology, 2006, v. 80, no. 15, p. 7450-7458, described that metal chelators (EDTA and 1,10-phenanthroline) were effective protease inhibitors assessed against NS2/3 autocleavage. There is also evidence that 1,10-phenanthroline acted in this case precisely through chelation of zinc.

In the article Sperandio D., Gangloff AR, Litvak J. Goldsmith R., Bioorg. Med. Chem. Lett., 2002, v.12, No. 21, 3129-3133, describes the screening of a group of bisbenzimidazoles in order to search for inhibitors of the serine protease NS3/NS4A of the hepatitis C virus, which also led to the identification of a corresponding powerful Zn ²⁺ -dependent inhibitor.

When developing new approaches to the treatment of hepatitis C, another attractive target is the RNA-dependent RNA polymerase of the hepatitis C virus (viral protein NS5B), which has a zinc-binding site in its structure [Timothy L. Tellinghuisen, Matthew S. Paulson, Charles M. Rice, J. Virology, 2006, v. 80, no. 15, p. 7450-7458].

Normally, the liver cell does not contain proteins with similar activity [Kozlov M.V., Polyakov K.M., Ivanov A.V., Biochemistry, 2006, v. 71, no. 9, pp. 1253-12594].

Currently known inhibitors of the RNA-dependent RNA polymerase of the hepatitis C virus can be divided into two main classes: nucleoside derivatives and non-nucleoside inhibitors of various natures [Maria Bretner, Acta biochemica polonica, 2005, v.52, No. 1, p. 57-70]. In addition, inhibition of the activity of this enzyme by pyrogallol derivatives was found. It is noteworthy that the mechanism of inhibition by pyrogallol derivatives is believed to be the chelation of magnesium cations taking part in the catalytic act at the stage of transfer of the phosphoryl residue [Kozlov M.V., Polyakov K.M., Ivanov A.V., Biochemistry, 2006, 71, No. 9, pp. 1253-12594].

Diseases caused by herpes viruses are widespread. Thus, several human herpesviruses are known - HCA simplex virus 1 and 2 (HSV-1 and HSV-2), cytomegalovirus (CMV), varicella zoster virus, Epstein-Barr virus. The destructive effects that this has on the central nervous system cause diseases such as encephalitis and meningitis. There may be interest in studying the effect of zinc chelating compounds, for example, diethylenetriamine pentaacetic acid, on the inhibition of human cytomegalovirus replication in vitro [Kanekiyo M., Itoh N., Mano M., Antiviral Res., 2000, v. 47, p. 207-214].

However, it should be noted that herpesviruses, like the above viruses, have proteins containing a zinc finger motif. Chemical changes in the “zinc finger” can lead to the release of zinc and changes in the structure of the functioning of viral proteins [Yan Chen, Christine M. Livingston, Stacy D. Carrington-Lawrence, J. of Virology, 2007, v.81, No. 16, p . 8742-8751].

“Zinc fingers” can serve as a target for new generation antiviral drugs. Several such compounds have already been identified. However, at present their effectiveness can only be judged based on in vitro studies.

The above information allows us to assert the important role of metal ions both in the normal functioning of cells and the whole organism, and in the development of pathologies. The ability of some compounds to chelate metal ions can serve as the basis for the creation of drugs capable of treating a variety of diseases, in particular viral ones.

In the article Megan Whitnall, Jonathan Howard, Prem Ponka, “A class of iron chelators with a wide spectrum of potent antitumor activity that overcomes resistance to chemotherapeutics,” PNAS, 2006, v. 103, no. 40, p. 14901-14906, effective iron chelators have been disclosed, which demonstrate high antiproliferative and antitumor activity, comparable to the activity of known cytostatics, and are promising for clinical studies.

In the article Kik K., Szmigiero L., “Dexrazoxane (ICRF-187) - a cardioprotectant and modulator of some anticancer drugs”, Postepy Hig Med Dosw Online, 2006, v. 60, p. 584-590, it is indicated that some iron chelators can be used as “assistants” in anticancer therapy, as they have a cardioprotective effect.

Chelation of copper ions inhibits angiogenesis and reduces tumor growth [Yu Yu, Jacky Wong, David B. Loveioy, “Chelators at the cancer coal face: Desferrioxamine to Triapine and Beyond,” Clin. Cancer Res., 2006, v. 12, p. 6876-6883].

The zinc chelator clioquinol, by binding Zn ²⁺ ions, causes apoptosis of human cancer cells [Haijun Yu, Yunfeng Zhou, Stuart E. Lind, “Clioquinol targets zinc to lysosomes in human cancer cells,” Biochem. J., 2009, v. 417, p. 133-139].

As aldehyde dehydrogenase inhibitors, chelators are used to treat alcoholism [Shian S.G., Kao Y.R., Wu F.Y., Wu C.W., “Inhibition of invasion and angiogenesis by zinc-chelating agent disulfiram,” Mol. Pharmacol., 2003, v. 64(5), p. 1076-84], as well as in alcoholic cirrhosis of the liver, iron excess anemia, porphyria tarda [Schroterova L., Kaiserova H., Baliharova V., “The effect of new lipophilic chelators on the activities of cytosolic reductases and P450 cytochromes involved in the metabolism of antracyclin as antibiotics: in vitro studies”, Physiol Res., 2004, v. 53(6), p. 683-691].

The activity of some antioxidants is due to the chelation of transition metal ions (Fe, Cu), which is accompanied by a decrease in metal-dependent lipid peroxidation [Babizhayev M.A., Seguin Marie-C., Gueynej J., Evstigneev R.P., Ageyeva E.A., Zheltuchina G.A., “L-Carnosine(- alanyl-L-histidine) and carcinine f-alanylhistamine) act as natural antioxidants with hydroxyl-radical-scavenging and lipid-peroxidase activities”, Biochem. J., 1994, v. 304, p. 509-516].

The use of antioxidants can promote the resorption of cataracts, eliminate retinal diseases and reduce the need for insulin in diabetics, eliminate skin pigmentation, and also help eliminate the effects of stroke. Chelation is also useful in the treatment of inflammatory diseases such as osteoarthritis, rheumatoid arthritis. [Zelenin K.N., “Complexons in medicine”, Soros Educational Journal, 2001, vol. 7, no. 1, pp. 45-50].

Chelators can be used in medicine as complexons for transporting and easily removing arsenic, mercury, antimony, cobalt, zinc, chromium, nickel from the body [Zholnin A.V., “Complex compounds”, Chelyabinsk: ChSMA, 2000, p. 28 ].

Inhibition of botulinum toxin by chelation of zinc ions is known [Anne C., Blommaert A., “Thioderivede disulfides as potent inhibitors of botulinum neurotoxin B: implications of zinc interaction,” Bioorg. Med. Chem., 2003, v. 11(21), p. 4655-60], in addition, chelation protects against gas gangrene [Zelenin K.N., “Complexons in medicine”, Soros Educational Journal, 2001, vol. 7, no. 1, pp. 45-50].

Chelation therapy is useful in the treatment of neurodegenerative diseases, in particular Alzheimer's disease, helping to improve memory [Bossy-Wetzel E., Schwarzenbacher R., Lipton S.A., “Molecular pathways to neurodegeneration”, Nat. Med, 2004, v. 10, p. 2-9]; Parkinson's disease [Kevin J. Barnham, Colin L. Masters, Ashley I. Bush, “Neurodegenerative diseases and oxidative stress,” Nature Reviews Drug Discovery, 2004, v. 3, p. 205-214]; Wilson's disease [Yu Yu, Jacky Wong, David B. Lovejoy, “Chelators at the Cancer Coalface: Desferrioxamine to Triapine and Beyond,” Clin. Cancer Res.? 2006, v. 12, p. 6876-6883]; Huntington's disease [Whitnall M., Richardson D.R., “Iron: a new target for pharmacological interference in neurodegenerative diseases”, Semin Pediatr Neurol, 2006, v. 13, p. 186-197]; amyotrophic lateral sclerosis [Kevin J. Burnham, Colin L. Masters, Ashley I. Bush, “Neurodegenerative diseases and oxidative stress”, Nature Reviews Drug Discovery, 2004, v. 4] and prion diseases [Daniel L. Cox, Jianping Pan, Rajiv R.P. Singh, “A Mechanism for Copper Inhibition of Infection Prion Conversion”, Biophysical Journal, 2006, v. 91, L11-L13]. Chelators prevent cancer [Megan Whitnall, with a wide spectrum of potent antitumor activity that overcomes resistance to chemotherapeutics”, PNAS, 2006, v. 103, no. 40, p. 14901-14906].

A significant place among the known chelators is occupied by derivatives of heterocyclic compounds, for example, imidazole, containing imido and amido groups.

In the article M.A. Podyminogin, V.V. Vlassov, “Synthesis of RNA-cleaving molecules mimicking ribonuclease A active center. Design and cleavage of tRNA transcrints”, Nucleic Acids Research, 1993, v. 21, No. 25, pp. 5950-5956, a bishistamine derivative of glutaric acid is described, which can serve as a model of the active site of nucleases and exhibits weak activity in the cleavage of RNA molecules.

In Elfriede Schuhmann et al., “Bis[platinum(II)] and Bis[Palladium (II)] complexes of α,ώ-Dicarboxylic Acid Bis(1,2,4-triaminobutane-N ⁴ )-Amides”, Inorg . Chem., 1995, v. 34, p. 2316-2322, bishistamine derivatives of glutaric and adipic acids are described, which are intermediates for the synthesis of complexes with platinum and palladium:

n=3-6.8

M=Pt, Pd

A method for the synthesis of N ¹ ,N ¹ -glutarylbis(histamine), including the interaction of histamine dihydrochloride and glutaric acid dichloride in dimethylformamide in the presence of a 4-fold excess of triethylamine, is described in the article by Elfriede Schuhmann et al., “Bis[platinum(II)] and Bis[ Palladium (II)] complexes of α,ώ-Dicarboxylic Acid Bis(1,2,4-triaminobutane-N ⁴ )-Amides”, Inorg. Chem., 1995, v. 34, p. 2316-2322.

The authors of the present invention discovered for the first time that the bishistamine derivative of glutaric acid, namely N ¹ ,N ¹ -glutaryl bis(histamine), is capable of forming complexes with metal ions.

Thus, the purpose of the present invention is to obtain biocompatible heterocyclic chelators of metal ions and their use as a medicine for the treatment and/or prevention of various diseases, using the ability of the claimed compounds to chelate metal ions.

The objective of the invention is also to develop simple methods for preparing such compounds using readily available reagents.

Brief description of the invention

The present invention relates to dicarboxylic acid bisamide derivatives of general formula I:

wherein

R ₁ represents a 5-membered unsaturated heterocyclic group containing from 1 to 2 heteroatoms selected from N and/or S, optionally fused with a 6-membered unsaturated cyclic group;

R ₂ represents a -C(O)-R ₃ -C(O)- group, where R ₃ represents a -(CH ₂ ) _n - group, optionally substituted with one or two C ₁ -C ₆ alkyls, or phenyl,

n is an integer from 0 to 4;

or pharmaceutically acceptable salts thereof.

The present invention also relates to derivatives of dicarboxylic acid bisamides of general formula I, which have the ability to chelate metal ions (Zn, Cu, Fe, Mg, Ca, etc.); as well as their use as a means for the prevention and/or treatment of cardiovascular, viral, oncological, neurodegenerative, inflammatory diseases, diabetes, gerontological diseases, as well as diseases caused by microorganism toxins, as well as alcoholism, alcoholic cirrhosis of the liver, anemia, late porphyria, poisoning with transition metal salts.

The present invention also relates to methods for preparing compounds of general formula I, including:

the reaction of a dicarboxylic acid and the corresponding amine upon heating; or

reacting the dicarboxylic acid with N-hydroxysuccinimide in the presence of N,N'-dicyclohexylcarbodiimide to produce the corresponding bis-N-hydroxysuccinimide ester, which is condensed with the amine; or

reacting the dicarboxylic acid diester with hydrazine hydrate, treating the resulting dihydrazide with sodium nitrite to obtain the corresponding azide, which is condensed with the amine; or

heating a solution of the imide formed from the dicarboxylic acid and the corresponding amine in an organic solvent; or

interaction of the corresponding amine and dicarboxylic acid in a 2:1 molar ratio in the presence of a condensing agent.

The proposed methods for preparing heterocyclic bis-derivatives of dicarboxylic acids of general formula I are simple to implement, proceed under fairly mild conditions, without the formation of by-products, are technologically advanced, and allow one to obtain target products with good yield (up to 82%) and a high degree of purity.

Detailed description of the invention

Preferred compounds of the present invention are those of general formula I:

,

where R ₁ represents a group selected from:

,

R ₂ represents a group selected from: -C(O)-(CH ₂ ) ₀ -C(O)-, -C(O)-(CH ₂ ) ₁ -C(O)-, -C(O) -(CH ₂ ) ₂ -C(O)-, -C(O)-(CH ₂ ) ₃ -C(O)-, -C(O)-(CH ₂ ) ₄ -C(O)-, - C(O)-CH ₂ -CH(CH ₃ )-CH ₂ -C(O)-, -C(O)-CH ₂ -C(CH ₃ ) ₂ -CH ₂ -C(O)-, or group

The most preferred compounds of the present invention are those shown in Table 1.

As pharmaceutically acceptable salts of the compounds of the present invention, addition salts of organic acids (for example, formate, acetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.), addition salts of inorganic acids (for example, hydrochloride, hydrobromide, sulfate, phosphate, etc.), salts with amino acids (for example, aspartic acid salt, glutamic acid salt, etc.), preferably hydrochlorides and acetates.

The most preferred known compounds that can be used in the pharmaceutical composition and treatment method of the present invention are the glutarimidide derivatives shown in Table 2.

The compounds of the present invention can be prepared by a process involving the condensation of a dicarboxylic acid of general formula II:

R ₄ OC(O)-R ₃ -C(O)-OR ₄ ,

where R ₃ represents a -(CH ₂ ) _n - group, optionally substituted with one or two C ₁ -C ₆ alkyls, or phenyl,

n is an integer from 0 to 4,

R ₄ represents hydrogen, C ₁ -C ₆ alkyl,

and an amine of general formula III:

NH ₂ -(CH ₂ ) ₂ -R ₁ ,

where R ₁ represents a 5-membered unsaturated heterocyclic group containing from 1 to 2 heteroatoms selected from N and/or S, optionally fused with a 6-membered unsaturated cyclic group;

by heating, optionally in the presence of a solvent.

It is preferable to use dimethyl ether and heat to a temperature of 150-170ºC, even more preferably to carry out condensation at boiling.

Diglyme or alcohols can be used as solvents, most preferably isoamyl alcohol.

Another method for preparing dicarboxylic acid bisamide derivatives of general formula I is a method involving the interaction of a dicarboxylic acid of general formula II:

R ₄ OC(O)-R ₃ -C(O)-OR ₄ ,

where R ₃ represents a -(CH ₂ ) _n - group, optionally substituted with one or two C ₁ -C ₆ alkyls, or phenyl,

n is an integer from 0 to 4,

R ₄ represents hydrogen,

with N-hydroxysuccinimide in the presence of N,N'-dicyclohexylcarbodiimide in N,N-dimethylformamide to give the corresponding bis-N-hydroxysuccinimide ester, general formula IV:

,

which is condensed with an amine of general formula III:

NH ₂ -(CH ₂ ) ₂ -R ₁ ,

where R ₁ represents a 5-membered unsaturated heterocyclic group containing from 1 to 2 heteroatoms selected from N and/or S, optionally fused with a 6-membered unsaturated cyclic group.

Cooling to a temperature of 0-5ºC is preferable.

Another method for preparing dicarboxylic acid bisamide derivatives of general formula I is a method involving the interaction of a dicarboxylic acid ester of general formula II:

R ₄ OC(O)-R ₃ -C(O)-OR ₄ ,

where R ₃ represents a -(CH ₂ ) _n - group, optionally substituted with one or two C ₁ -C ₆ alkyls, or phenyl,

n is an integer from 0 to 4,

R ₄ represents C ₁ -C ₆ alkyl,

with hydrazine hydrate in an organic solvent to obtain a bishydrazide of general formula V:

H ₂ N-NH-C(O)-R ₃ -C(O)-NH-NH ₂ ,

treatment of bishydrazide with sodium nitrite in an acidic environment at a temperature of about 0ºC to obtain bisazide of general formula VI:

N ^- -N ⁺ =NC(O)-R ₃ -C(O)-N=N ⁺ -N ^- ,

condensation of a bisazide with an amine of general formula III:

NH ₂ -(CH ₂ ) ₂ -R ₁ ,

where R ₁ represents a 5-membered unsaturated heterocyclic group containing from 1 to 2 heteroatoms selected from N and/or S, optionally fused with a 6-membered unsaturated cyclic group, in an organic solvent.

Preferably, alcohols are used as the organic solvent, most preferably isopropanol. The method is simple, but is applicable if the number of methylene units in the original dicarboxylic acid is greater than or equal to three, since the bisazides obtained during the synthesis for acids with a smaller number of methylene groups are unstable.

Dicarboxylic acid bisamidide derivatives of general formula I can also be prepared by a process involving condensation of an imide of general formula VII:

,

where R ₃ represents a -(CH ₂ ) _n - group, optionally substituted with one or two C ₁ -C ₆ alkyls,

n is an integer from 0 to 4,

with an equimolar amount of amine of general formula III:

NH ₂ -(CH ₂ ) ₂ -R ₁ ,

where R ₁ represents a 5-membered unsaturated heterocyclic group containing from 1 to 2 heteroatoms selected from N and/or S, optionally fused with a 6-membered unsaturated cyclic group,

in an organic solvent when heated.

Preferably, alcohols are used as an organic solvent, most preferably isopropanol, and the condensation is carried out at boiling.

Another method of the present invention is a method for the preparation of bisamide derivatives of dicarboxylic acids of general formula I, including the reaction of a dicarboxylic acid of general formula II:

R ₄ OC(O)-R ₃ -C(O)-OR ₄ ,

where R ₃ represents a -(CH ₂ ) _n - group, optionally substituted with one or two C ₁ -C ₆ alkyls, or phenyl,

n is an integer from 0 to 4,

R ₄ represents hydrogen,

and an amine of general formula III:

NH ₂ -(CH ₂ ) ₂ -R ₁ ,

where R ₁ represents a 5-membered unsaturated heterocyclic group containing from 1 to 2 heteroatoms selected from N and/or S, optionally fused with a 6-membered unsaturated cyclic group;

at a molar ratio of 1:2-2.5 in a solution of tetrahydrofuran in the presence of a condensing agent, preferably carbonyldiimidazole.

The present invention also relates to a medicinal product, a pharmaceutical composition containing derivatives of bisamides of dicarboxylic acids of general formula I, and a method including the administration of derivatives of bisamides of dicarboxylic acids of general formula I, for the prevention and/or treatment of viral diseases in humans and animals, including diseases caused by hepatitis C virus, human papillomavirus, HIV or oncogenic RNA viruses such as leukemia virus; cardiovascular diseases, including diseases caused by cardiotoxicity of cytostatics, cholesterol deposition, high blood pressure; diseases associated with metal-dependent reactions of free radical oxidation, including gerontological diseases such as cataracts, retinal diseases, skin pigmentation; consequences of stroke; atherosclerosis; inflammatory diseases such as osteoarthritis, rheumatoid arthritis;

diabetes and its vascular complications; neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, Wilson's disease, Huntington's disease, amyotrophic lateral sclerosis, prion diseases;

oncological diseases; diseases caused by microorganism toxins, in particular botulism or gas gangrene;

alcoholism and alcoholic cirrhosis of the liver; iron excess anemia, porphyria tarda; poisoning with transition metal salts.

The compounds of the present invention are administered in an effective amount that provides the desired therapeutic result.

The compounds of general formula (I) can be administered orally, topically, parenterally, intranasally, inhaled and rectally in unit dosage forms containing non-toxic pharmaceutically acceptable carriers. As used herein, the term “parenteral administration” means subcutaneous, intravenous, intramuscular or intrathoracic injections or infusions.

The compounds of the present invention can be administered to a patient in doses ranging from 0.1 to 100 mg/kg body weight per day, preferably in doses from 0.25 to 25 mg/kg one or more times per day.

It should be noted that the specific dose for each individual patient will depend on many factors, including the potency of the compound used, age, body weight, gender, the patient's general health and diet, the time and method of administration of the drug, the rate of its elimination from the patient. body, the specific combination of drugs used, and the severity of the disease in the individual being treated.

The pharmaceutical compositions of the present invention contain a compound of general formula (I) in an amount effective to achieve the desired result, and can be administered in unit dosage forms (for example, solid, semi-solid or liquid form) containing the compounds of the present invention as the active ingredient mixed with a carrier or excipient suitable for intramuscular, intravenous, oral, sublingual, inhalation, intranasal and intrarectal administration. The active ingredient may be formulated together with commonly used non-toxic pharmaceutically acceptable carriers suitable for the preparation of solutions, tablets, pills, capsules, dragees, emulsions, suspensions, ointments, gels and any other dosage forms.

Various substances can be used as fillers, such as saccharides, for example glucose, lactose or sucrose, mannitol or sorbitol, cellulose derivatives and/or calcium phosphates, for example tricalcium phosphate or calcium hydrogen phosphate; components such as starch paste, for example, corn, wheat, rice, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone. Disintegrants such as the aforementioned starches and carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof such as sodium alginate may be used if necessary.

Optional additives may be used such as flow control agents and lubricants such as silica, talc, stearic acid and its salts such as magnesium stearate or calcium stearate, and/or propylene glycol.

The dragee core is usually coated with a layer that is resistant to gastric juice. For this purpose, concentrated solutions of saccharides can be used, which may optionally contain gum arabic, talc, polyvinylpyrrolidone, polyethylene glycol and/or titanium dioxide, and suitable organic solvents or mixtures thereof.

Stabilizers, thickeners, dyes and fragrances can also be used as additives.

Hydrocarbon ointment bases, such as white and yellow petroleum jelly (Vaselinum album, Vaselinum flavum), petroleum jelly (Oleum Vaselini), white and liquid ointment (Unguentum album, Unguentum flavum), can be used as an ointment base, and as additives to give denser consistency - such as paraffin and wax; absorbent ointment bases such as hydrophilic petroleum jelly (Vaselinum hydrophylicum), lanolin (Lanolinum), cold cream (Unguentum leniens); water-rinse ointment bases such as hydrophilic ointment (Unguentum hydrophylum); water-soluble ointment bases, such as polyethylene glycol ointment (Unguentum Glycolis Polyaethyleni), bentonite bases and others.

Methylcellulose, sodium salt of carboxymethylcellulose, hydroxypropylcellulose, polyethylene glycol or polyethylene oxide, carbopol can be used as a base for gels.

As a base for the suppository, bases that are insoluble in water, such as cocoa butter, can be used; bases soluble in water or miscible with water, such as gelatin-glycerin or polyethylene oxide; combined bases - soap and glycerin.

When preparing a unit dosage form, the amount of active ingredient used in combination with the carrier may vary depending on the recipient being treated and the particular route of administration of the drug.

For example, when using the compounds of the present invention in the form of injection solutions, the content of the active agent in them is up to 5% by weight. As diluents, 0.9% sodium chloride solution, distilled water, novocaine injection solution, Ringer's solution, glucose solution, and specific additives for dissolution can be used. When the compounds of the present invention are administered into the body in the form of tablets and suppositories, their amount is up to 200 mg per standard dosage form.

The dosage forms of the present invention are prepared by standard techniques such as, for example, mixing, granulating, dragee forming, dissolution and lyophilization processes.

A detailed description of the compounds of the present invention, their preparation and activity studies is presented in the following examples, intended to illustrate preferred embodiments of the invention and not to limit its scope.

Examples of the synthesis of glutarimidide derivatives of general formula I

Means and methods

The identity of the resulting compounds is checked by TLC on Kieselgel 60 F254 plates (Merck, Germany) in a solvent system: pyridine-acetic acid-water (20:6:11) - system A, A:ethyl acetate 3:1 (1 ), chloroform-methanol 9:1 (2).

Electrophoresis on paper (electrophoresis paper of the Kondopoga Pulp and Paper Mill, 120×320 mm) is carried out in a buffer with pH 5.1 of the composition pyridine-acetic acid-water 12:10:1000 in a chamber (150×320×150 mm) with a gradient of 15 V/ cm for 1.5 hours.

Chromatograms and electropherograms are developed with chloro-tetramethylbenzidine reagent and Pauli reagent.

The melting point is determined using a PTP device (laboratory instrument plant, Russia, Klin).

FTIR spectra are recorded in KBr tablets on a Magna 750 instrument (Nicolet (USA)).

¹ H-NMR spectra are recorded on an AMX-400 (Germany), Bruker DPX-400 (Germany) instrument.

High-resolution mass spectra are obtained on a time-of-flight mass spectrometer using matrix laser desorption ionization using 2,5-dihydroxybenzoic acid as a matrix, on an Ultraflex device (Bruker, Germany).

LC/MS system for the analysis of multicomponent mixtures Shimadzu Analytical HPLC SCL10Avp, mass spectrometer PE SCIEX API 165 (150), (Canada).

Analytical reverse-phase HPLC was carried out on a device: HPLC Shimadzu chromatograph under conditions: column Luna C18 (2) 100 A, 250×4.6 mm (ser. 599779-23), elution gradient in a phosphate buffer solution pH 3.0: methanol (conditions A). The device uses a Beckman (USA) “System Gold” chromatograph with a UV detector (λ=220 nm), a “Gemini” C-18 column, 150×2.0 mm Phenomenex, a gradient mobile phase in 0.05 M phosphate buffer (pH 3.0) 90% MeCN in water, elution rate 0.25 ml/min, injection of 10 μl (conditions B).

Example 1

Bis-1,5-(N ^β -histaminyl)glutaric acid; (N,N'-bis-[2-(1H-imidazol-4-yl)ethyl]pentanediamide (compound 11)

To 5 g (0.031 mol) of glutaric acid dimethyl ester, add 8 g (0.072 mol) of histamine and heat at 170ºC for 3.5-4 hours until the release of methyl alcohol vapors stops. The completeness of the reaction is monitored by TLC or electrophoresis. Then the reaction mass is suspended in isopropyl alcohol and left for 24 hours at +4ºС. The product is separated, washed with isopropyl alcohol, and dried. Yield 6.8 g (69%). R_f 0.42 (1). E⁺49 mm. T.pl. 166-168ºС. LC/MS, individual peak, retention time 0.3 min [M+H]⁺=319. HPLC under conditions A, individual peak, retention time 5.58 min. Range¹H-NMR (400.13 MHz, DMSO-d₆, δ, ppm, J/Hz): 1.70 (pent., 2H, CH₂C H ₂ CH₂, J=7.3 Hz), 2.03 (t, 4H, C H ₂CH ₂ C H ₂, J=7.3 Hz), 2.61 (t, 4H, CC H ₂ CH₂N, J=7.5 Hz), 3.26 (square, 4H, CCH ₂ C H ₂N, J=7.5 Hz), 6.76 (broad s, 2H, CC H ), 7.50 (s, 2H, NC H N ), 7.94 (lat, 2H, N H ). IR-Fourier spectrum (in the table KBr, ν, cm^-1): 1651 (ν C=O amide I), 1580 (δ NH amide II), 1425 (-CH ₂ -SO-). Found, %: C 56.40, H 6.86, N 26.31. WITH₁₅N₂₂N₆O₂. Calculated, %: C 56.59, H 6.96, N 26.40.

Example 2

Bis-1,4-(N ^β -histaminyl)succinic acid; (N,N'-bis-[2-(1H-imidazol-4-yl)ethyl]butanediamide) (compound 3)

12.2 g (0.11 mol) of histamine are added to 5.5 g (0.046 mol) of succinic acid and heated at 170ºC for 3.5-4 hours until the release of water vapor stops. The completeness of the reaction is monitored by TLC or electrophoresis. Then the reaction mass is suspended in 50 ml of water and left for 24 hours at +4ºС. The product is separated, washed with water, and dried. Yield 10.7 g (76%). R _f 0.51 (1). E ⁺ 51 mm. T.pl. 230-231ºС. [M+H] ⁺ 304.95. ¹ H-NMR spectrum (D ₂ O): δ, ppm: 2.48 (s, 4H, CH ₂ -Suc), 2.80-2.84 (t, 4H, β-CH ₂ -HA ), 3.44-3.48 (t, 4H, α-CH ₂ -HA), 6.97 (s, 2H, 5-CH-Im), 7.78 (s, 2H, 2-CH-Im ). IR-Fourier spectrum (in the table KBr, ν, cm ^-1 ): 1634 (ν C=O amide I), 1583 (δ NH amide II), 1429 (- CH ₂ -CO-). Found, %: C 55.40, H 6.66, N 27.31. C ₁₄ H ₂₀ N ₆ O ₂ . Calculated, %: C 55.25, H 6.62, N 27.61.

Example 3

Bis-1,3-(N ^β -histaminyl)malonic acid; (N,N'-bis-[2-(1H-imidazol-4-yl)ethyl]propanediamide) (compound 2)

To 6.4 g (0.039 mol) of malonic acid diethyl ester, add 9.0 g (0.081 mol) of histamine and heat at 170ºC for 2.5-3 hours until the release of ethyl alcohol vapor stops. The completeness of the reaction is monitored by TLC or electrophoresis. Then the reaction mass is suspended in 20 ml of water and left for 72 hours at +4ºС. The product is separated, washed with isopropyl alcohol, and dried. Yield 5.0 g (44%). R _f 0.41 (1). E ⁺ 57 mm. T.pl. 187-188ºС. ¹ H-NMR spectrum (D ₂ O): δ, ppm: 2.74-2.78 (t, 4H, β-CH ₂ -HA), 3.15 (s, 2H, CH ₂ -Mal ), 3.41-3.44 (t, 4H, α-CH ₂ -HA), 6.78 (s, 2H, 5-CH-Im), 7.66 (s, 2H, 2-CH-Im ). IR-Fourier spectrum (in the table KBr, ν, cm ^-1 ): 1643 (ν C=O amide I), 1574 (δ NH amide II), 1426 (- CH ₂ -CO-). Found, %: C 53.24, H 6.51, N 28.56. C ₁₃ H ₁₈ N ₆ O ₂ . Calculated, %: C 53.78, H 6.25, N 28.95.

Example 4

Bis-1,2-(N ^β -histaminyl)oxalic acid; (N,N'-bis-[2-(1H-imidazol-4-yl)ethyl]ethanediamide) (compound 1)

To a solution of 7.3 g (0.05 mol) of oxalic acid diethyl ester in 35 ml of isoamyl alcohol, add 12.2 g (0.11 mol) of histamine and boil for 4 hours. The completeness of the reaction is monitored by TLC or electrophoresis. The reaction mixture is left for 16 hours at +4ºС. The precipitate is separated, washed with isopropyl alcohol, and dried. Yield 10 g (72%). R _f 0.55 (1). E ⁺ 55 mm. T.pl. 235-236ºС. ¹ H-NMR spectrum (DMSO): δ, ppm: 2.66-2.72 (t, 4H, β-CH ₂ -HA), 3.33-3.41 (m, 4H, α- CH ₂ -HA), 6.81 (s, 2H, 5-CH-Im), 7.54 (s, 2H, 2-CH-Im), 8.78-8.84 (t, 2H, NH - CO). IR-Fourier spectrum (in the table KBr, ν, cm ^-1 ): 1651 (ν C=O amide I), 1566 (δ NH amide II), 1425 (- CH ₂ -CO-). Found, %: C 52.24, H 5.51, N 29.97. C ₁₂ H ₁₆ N ₆ O ₂ . Calculated, %: C 52.17, H 5.84, N 30.42.

Example 5

Bis-1,3-(N ^β -histaminyl)isophthalic acid (compound 6)

5 g (30 mmol) of isophthalic acid and 7.93 g (69 mmol) of N-hydroxysuccinimide are dissolved in 30 ml of dimethylforamide. To the resulting solution, cooled to 5ºC, add a cooled solution of 14.24 g (69 mmol) of N,N'-dicyclohexylcarbodiimide in 10 ml of dimethylforamide, the reaction mixture is left overnight at 5ºC. The urea is separated, washed with 15 ml of dimethylformamide and dried. Based on the mass of the urea obtained, the reaction yield is considered to be 80%. A solution of N-hydroxysuccinimide diester of isophthalic acid is cooled to 5ºC, after which a melt of 6.84 g (61.6 mmol) of histamine is added to it in portions. The reaction mixture is left for 2 hours. The solvent is removed in vacuo. The residue, a yellow oil, was dissolved in 120 ml of water and passed through an Amberlite IRA-96 column (27x115). Fractions containing the product are combined. The precipitated crystals of the target product are separated. The resulting technical product is recrystallized from a mixture of 35 ml of isopropanol and 10 ml of water. The product crystals are filtered, washed with water (3×30 ml) and isopropanol (2×13 ml), and dried. The target product is obtained in the form of white crystals. Yield 1.55 g (18% based on histamine, or 14% based on isophthalic acid). R _f 0.42 (1). T.pl. 218-219ºС. E ⁺ 40 mm. Macc-spectrum: [M+1] ⁺ 353 _. ¹ H-NMR spectrum: (300 MHz, DMSO-d ₆ ): δ, ppm: 2.74-2.79 (t, 4H, β-CH ₂ -Ha), 3.46-3.52 (t , 4H, α-CH ₂ -Ha), 6.81 (s, 2H, 5-CH-Im), 7.52-7.56 (m, 3H, ArH+2-CH-Im), 7.92 -7.95 (d, 2H, ArH), 8.28 (s, 1H, ArH), 8.65 (brs, 2H, -C(O)-NH-), 11.80 (brs , 2H, -NH-). HPLC under conditions B, individual peak, recovery time 12.5 min.

Example 6

Bis-1,5-(N ^β -histaminyl)glutaric acid; (N,N'-bis-[2-(1H-imidazol-4-yl)ethyl]pentanediamide) (compound 11)

To a solution of 13.2 g (0.10 mol) of glutaric acid in 50 ml of methanol, 8 ml of PCl ₃ is added in portions while cooling and stirring. The solvent from the reaction mixture is removed in vacuo. The resulting residue is distilled in vacuum. 14.7 g (92%) of glutaric acid dimethyl ester are obtained, bp. 110-112ºС.

7.5 ml of hydrazine hydrate is added to 20 ml of isopropyl alcohol, heated to a boil and 8 g (0.05 mol) of glutaric acid dimethyl ester are added dropwise, the reaction mixture is left at +20ºC for 16 hours. The precipitated product is separated, washed with isopropyl alcohol. alcohol. Dry. 7.3 g (91%) of glutaric acid dihydrazide are obtained, m.p. 210-212ºС.

To a solution of 1.61 g (0.010 mol) of glutaric acid dihydrazide in a mixture of 20 g of ice with 2.5 ml of hydrochloric acid and 10 ml of pre-cooled chloroform, 1.45 g (0.021 mol) of sodium nitrite is added in portions with stirring. The chloroform layer is separated, washed with 10 ml of water cooled to +4ºС and added to a cooled solution of 2.6 g (0.023 mol) of histamine in 5 ml of isopropanol. The reaction mixture is left at +20ºС for 2 hours, then the solvent is removed in vacuum. The resulting residue is dissolved in 20 ml of water and passed through a column with 50 ml of KU-2-20 ion exchange resin in H ⁺ form. The resin is washed with a 0.5% ammonia solution, collecting fractions of the histamine-free product. The eluates are combined, the solvent is removed in vacuo, the resulting residue is recrystallized from 2 ml of isopropanol and dried. 1.8 g (55%) are obtained. R _f 0.42 (1). E ⁺ 49 mm. T.pl. 166-168ºС. ¹ H-NMR spectrum (D ₂ O): δ, ppm: 1.73-1.78 (m, 2H, β-CH ₂ -Glt), 2.10-2.15 (t, 4H, α-CH ₂ -Glt), 2.78-2.82 (t, 4H, β-CH ₂ -HA), 3.43-3.48 (t, 4H, α-CH ₂ -HA), 6, 94 (s, 2H, 5-CH-Im), 7.69 (s, 2H, 2-CH-Im). IR-Fourier spectrum (in the table KBr, ν, cm ^-1 ): 1651 (ν C=O amide I), 1580 (δ NH amide II), 1425 (- CH ₂ -CO-). Found, %: C 56.40, H 6.86, N 26.31. C ₁₅ H ₂₂ N ₆ O ₂ . Calculated, %: C 56.59, H 6.96, N 26.40.

Example 7

Bis-1,5-(N ^β -histaminyl)adipic acid; (N,N'-bis-[2-(1H-imidazol-4-yl)ethyl]hexanediamide) (compound 12)

To a solution of 7.40 g (0.05 mol) of adipic acid in 25 ml of methanol, add 4 ml of phosphorus trichloride dropwise while cooling. The solvent from the reaction mixture is removed in vacuo, and the resulting residue is distilled in vacuo. 7.5 g (86%) of adipic acid dimethyl ester are obtained, bp. 115-117ºС.

To a mixture of 20 ml of isopropyl alcohol and 5 ml of hydrazine hydrate, heated to boiling, 7.5 g (0.04 mol) of adipic acid dimethyl ester is added dropwise, the reaction mixture is left at +20ºC for 16 hours. The precipitated product is separated and washed isopropyl alcohol, dry. 6.2 g (83%) of adipic acid dihydrazide are obtained, m.p. 182-182.5ºС.

To a solution of 2.2 g (0.013 mol) of adipic acid dihydrazide in a mixture of 30 g of ice with 3 ml of hydrochloric acid and 15 ml of cooled chloroform, 1.8 g (0.026 mol) of sodium nitrite is gradually sprinkled in portions with stirring. The chloroform layer is separated, washed with 10 ml of water cooled to +4ºС and added to a cooled solution of 3 g (0.027 mol) of histamine in 8 ml of isopropanol. The completeness of the reaction is monitored by TLC or electrophoresis. The reaction mixture is left for 24 hours at +4ºС, the precipitate is separated, washed with isopropyl alcohol, recrystallized from water, and dried. 2.8 g (65%) are obtained. R _f 0.48 (1). E ⁺ 45 mm. T.pl. 184-186ºС. [M+H] ⁺ 332.92. ¹ H-NMR spectrum (D ₂ O): δ, ppm: 1.37-1.41 (m, 4H, β-CH ₂ -Adip), 2.11-2.15 (m, 4H, α-CH ₂ -Adip), 2.74-2.78 (t, 4H, β-CH ₂ -HA), 3.40-3.44 (t, 4H, α-CH ₂ -HA), 6, 88 (s, 2H, 5-CH-Im), 7.63 (s, 2H, 2-CH-Im). IR-Fourier spectrum (in the table KBr, ν, cm ^-1 ): 1646 (ν C=O amide I), 1581 (δ NH amide II), 1425 (- CH ₂ -CO-). Found, %: C 57.45, H 7.15, N 24.97. C ₁₆ H ₂₄ N ₆ O ₂ . Calculated, %: C 57.81, H 7.28, N 25.28.

In accordance with the above procedure, the following compounds were obtained:

Connection number Structural formula Physico-chemical data Connection 8 Macc-spectrum: [M+1] ⁺ =353. HPLC: under conditions B, individual peak, recovery time 16.1 min. ¹ H-NMR spectrum (400 MHz, DMSO-d ₆ ): δ, ppm: 1.63-1.75 (2H, m, CH ₂ ), 1.96-2.11 (4H, t, J=7.46, 2CH ₂ ), 3.06-3.16 (4H, t, J=6.97, CH ₂ ), 3.35-3.46 (4H, dd, J1=6.61, J2=6.11, CH ₂ ), 7.66-7.74 (2H, d, J=3.18, Ar), 7.66-7.74 (2H, d, J=3.18, Ar ), 7.9-8.0 (2H, m, NH). Connection 10 LC/MS, individual peak, retention time 0.3 min [M+H]⁺=319. HPLC under conditions A, individual peak, retention time 4.12 min. Range¹H-NMR (400.13 MHz, DMSO-d₆, δ, ppm, J/Hz): 1.69 (pent., 2H, CH₂C H ₂ CH₂, J=7.3 Hz), 2.01 (t, 4H, C H ₂ CH ₂ C H ₂, J=7.3 Hz), 2.73 (t, 4H, CC H ₂ CH₂N, J=7.5 Hz), 3.31 (sq., 4H, CCH ₂ C H ₂ N, J=7.5 Hz), 6.87 (s, 4H, C H N ), 8.00 (lat, 2H, N H ).

Example 8

Bis-1,4-(N ^β -histaminyl)succinic acid; (N,N'-bis-[2-(1H-imidazol-4-yl)ethyl]butanediamide) (compound 3)

To a solution of 0.10 g (0.9 mmol) of histamine in 3 ml of isopropyl alcohol, add 0.15 g (0.78 mmol) of succinimidohistamine and heat to boiling for 2.5-3 hours. The completeness of the reaction is monitored by TLC or electrophoresis. Then the reaction mass is cooled to room temperature and left for 26 hours at +4ºС. The product is separated, washed with isopropyl alcohol, and dried. Yield 0.20 g (81%). R _f 0.44 (1). E ⁺ 51 mm. T.pl. 231ºС. [M] ⁺ 304.12. ¹ H-NMR spectrum (D ₂ O): δ, ppm: 2.48 (s, 4H, CH ₂ -Suc), 2.80-2.84 (t, 4H, β-CH ₂ -HA ), 3.44-3.48 (t, 4H, α-CH ₂ -HA), 6.97 (s, 2H, 5-CH-Im), 7.78 (s, 2H, 2-CH-Im ). IR-Fourier spectrum (in the table KBr, ν, cm ^-1 ): 1634 (ν C=O amide I), 1583 (δ NH amide II), 1429 (- CH ₂ -CO-). Found, %: C 6.57, N 27.18. C ₁₄ H ₂₀ N ₆ O ₂ . Calculated, %: C 55.25, H 6.62, N 27.61.

Example 9

Bis-1,5-(N-ethylamino-2-benzimidazolyl)glutaric acid; (N,N'-bis-[2-(1H-benzimidazol-2-yl)ethyl]pentanediamide) (compound 7)

To a solution of 8 g (0.06 mol) of glutaric acid in 500 ml of anhydrous tetrahydrofuran, add 23.4 g (0.145 mol) of carbonyldiimidazole, stir for 2 hours. Then 21.6 g (0.133 mol) of benzimidazolyl ethylamine are added to the reaction mixture. The resulting solution is stirred for 3 hours and left at room temperature. The product precipitate is separated, washed with water, and dried. 21 g (82.9%) are obtained. R _f 0.21 (2). T.pl. 170-170.5ºС. [M+1] ⁺ 419. ¹ H-NMR spectrum (DMSO): δ, ppm: 1.61-1.76 (m, 2H, CH ₂ ), 1.95-2.07 (t, 2H, CH ₂ ), 2.9-3.0 (t, 4H, 2CH ₂ ), 3.45-3.55 (q.v., 4H, 2CH ₂ ), 7.06-7.14 (m, 4H , Ar), 7.39-7.54 (m, 4H, Ar), 7.98-8.08 (t, 2H, Ar), 11.6-12.7 (s, 2H, NH). HPLC under conditions B, individual peak, recovery time 26.6 min.

In accordance with the above procedure, the following compounds were obtained:

Connection number Structural formula Physico-chemical data Connection 9 Macc-spectrum: [M+1] ⁺ =453. ¹ H-NMR spectrum: (400 MHz, DMSO-d ₆ ): δ, ppm: 1.65-1.77 (2H, m, CH ₂ ), 2.0-2.1 (2H, t , J=7.46, CH ₂ ), 3.16-3.26 (4H, t, J=6.97, 2CH ₂ ), 3.45-3.56 (4H, q., J1=6, 48, J2=5.87, 2CH ₂ ), 7.35-7.44 (1H, m, Ar), 7.44-7.52 (1H, m, Ar), 7.9-7.96 ( 2H, d, Ar, J=7.82), 7.9 6-8.02 (2H, t, NH, J=5.5), 8.02-8.07 (2H, d, Ar, J =7.82). HPLC under conditions B, individual peak, recovery time 18.7 min.

Study of the chelating ability of the described compounds

The proposed compounds of general formula I, as shown by further research, have the ability to complex or chelate metal ions. They have several functional groups that can act as electron donors during complex formation, for example, a carboxyl group, imido-, amido groups, and are ligands that specifically interact with metal ions, for example, with zinc, copper, iron, calcium, magnesium ions.

An important characteristic of the ability of a ligand to form complexes is the dissociation constants (pk _n ) of the compounds under study in an aqueous solution. The pk _n values are determined using the potentiometric titration method. Calculation of dissociation constants is carried out according to the Schwarzenbach method [Grinberg A.A., "Introduction to the chemistry of complex compounds", 4th ed., rev., Leningrad, Chemistry, 1971].

Methods of pH-potentiometric titration

Titration was carried out on a setup using a laboratory type ion meter I 120.1. The created galvanic cell consisted of two electrodes: an indicator glass and a silver chloride reference electrode. pH values were determined with an accuracy of ±0.2.

Initial 0.05 M solutions of metal ion salts M ^z+ were prepared from zinc nitrates (Zn(NO ₃ ) ₂ 6H ₂ O), calcium (Ca(NO ₃ ) ₂ 4H ₂ O), copper sulfate (CuSO ₄ 5H ₂ O), magnesium chloride (MgCl ₂ 6H ₂ O) and Mohr's salt ((NH ₄ ) ₂ Fe(SO ₄ ) ₂ 6H ₂ O). Standardization of the initial solutions of M ^z+ salts was carried out using complexometric titration with Trilon B (M ^z+ =Zn ²⁺ , Cu ²⁺ , Mg ²⁺ , Ca ²⁺ ), spectrophotometrically at 490 nm using o-phenanthroline (M ^z+ =Fe ²⁺ ).

Typical method for determining stepwise acid dissociation constants (k ₁ , . . ., k _n ) by pH-metric titration using the Schwarzenbach method

A weighed portion of the test compound (3⋅10^-4 mol) was dissolved in 15.0 ml of 0.5 M aqueous KNO solution₃. The resulting 0.02 M solution was titrated with vigorous stirring with a 0.05 M aqueous solution of NaOH or HCl. The amount (V, ml) of the consumed titrant and the pH of the solution were recorded at each titration point. The titration step was 0.5 ml, near the equivalence point - 0.2 ml. Titration was carried out until the pH of the solution was 11.0-11.5. From the titration curve pH=f(a), the pH value of the solution corresponding to a=0.5 and/or a=1.5 was determined, and the number of added g-equivalents of the titrant was determined per 1 mole of the titrated substance.

The results of determining the acid dissociation constants [pk _n ] using the Schwarzenbach method for the studied compounds are presented in Table 3.

Table 3
The pk values of the studied compounds in an aqueous solution, calculated using the Schwarzenbach method (C 0.02 M, ionic strength of the solution 0.5 (KNO ₃ ), 22ºС) Connection number рk ₁ рk ₂ 2 7.11 ⁽¹⁾ 11.27 ⁽²⁾ 3 6.88 ⁽¹⁾ 11.27 ⁽²⁾ eleven 7.16 ⁽¹⁾ 11.89 ⁽²⁾ 12 7.13 ⁽¹⁾ 11.95 ^{(2) (1)} - titration with aqueous 0.05 M HCl solution.
⁽²⁾ - titration with an aqueous 0.05 M NaOH solution.

Taking into account such parameters as the amount of the test substance required for potentiometric titration, the methodology for its implementation and the calculation of constants, the most suitable, informative and simple is the Bjerrum method [Galaktionov, M.A. Chistyakov, V.G. Sevastyanov et al., “Stepwise stability constants of complexes of group III B elements and lanthanides with acetylacetone in aqueous solution and the solubility products of their triacetylacetonates,” J. Phys. Khim., 2004, v. 78, no. 9, p. 1596-1604].

The method is based on the concept of a stepwise process of complex formation. The main assumption of Bjerrum's method is the assumption that only mononuclear complexes are formed. Schematically, the complex formation process can be described as follows:

M+L↔ML K ₁ =[ML]/[M][L]

ML _n-1 +L↔ML _n K _n =[ML _n ]/[ML _n-1 ][L]

According to Bjerrum's method, the compound under study is titrated twice with NaOH solution: in the absence and in the presence of a metal ion. Titration is carried out at a high constant concentration of an indifferent electrolyte, ensuring a constant ionic strength of the solution.

For these purposes, highly soluble salts are used, for example, potassium nitrate. Concentration stepwise stability constants ( _Kn ) are experimentally found. In addition, it is possible to obtain information about the composition of the resulting complexes, namely, the maximum possible number of attached ligands (n).

A typical method for determining stepwise stability constants of complexes by pH-metric titration using the Bjerrum method

A weighed portion of the test compound (3⋅10^-4 mol) was dissolved in 14.0 ml of 0.5 M aqueous KNO solution₃. With vigorous stirring, 1.0 ml (5⋅10^-4 mol) 0.05 M aqueous solution of salt M^z+. Systems with the relation M^z+To L 1:6. The pH values of the solution were recorded in the combined presence of the ligand and metal ions in the solution. Titration of the mixture was carried out with a 0.05 M aqueous solution of NaOH over a wide pH range (up to pH values of 11.0-11.5). The amount of titrant consumed at each titration point was recorded (V_NaOH, ml) and pH of the solution. At the moment of formation of a precipitate, turbidity of the solution or change in its color, the pH value, the nature of the change and the volume of the NaOH solution used for titration were recorded. The titration step is 0.25 ml. According to the Bjerrum curve=f(pA) for each half-integer value of the formation function=n-0.5 determined logK_n.

Currently, the Bjerrum method is often used to study complex formation. This method involves the determination of two parameters - the concentration of the free ligand under equilibrium conditions - [L ^- ] and the formation function , which is the average number of ligands included in the complex. Thus, at any moment of titration, using experimental data, it is possible to calculate [L ^- ] (namely, -log[L ^- ] = рА) and according to equation (1) - . When the ligand has several dissociating groups, then in equation (1) K _L corresponds to the value of the dissociation constant that has the largest pk value.

From the constructed Bjerrum curve (formation curve) =f(pA) according to equation (2) for each half-integer value of the formation function =n-0.5 find K _n . Thus, the stepwise stability constant will be equal to the inverse concentration of the free ligand at the point =n-0.5. Accordingly, the higher the K _n value, the more stable the complex compound (chelate) is.

Where And - total concentrations of ligand and metal present in the solution, mol/l;

[L ^- ] - concentration of free ligand ions, mol/l;

K _L is the acid dissociation constant of the ligand;

- volume of NaOH solution ( , mol/l) at any point in the titration, added to the titrated mixture of M ^z+ and L, l;

_nL , - amount of ligand substance and salt M ^z+ in the titrated solution, mol.

The logK ₁ values found using the Bjerrum method for complexes of the studied compounds with divalent metal ions are presented in Tables 4, 5. The choice of the M ^z+ to L ratio of 1:6 is due to the availability of data on the maximum possible coordination number for Zn(II) and Fe (II), equal to six, and for Cu(II) - equal to four [Claudia A. Blindauer, M. Tahir Razi, Simon Parsons, Peter J. Sadler, Polyhedron, 2006, v. 25, r. 513-520].

Table 4
The first stability constant of the 1:1 complexes of the studied compounds with divalent metal ions is logK ₁ , found using the Bjerrum method (aqueous solution, C _L =0.02 M, ratio M ^z+ To L 1:6, ionic strength 0.5 (KNO ₃ ), 22ºС) Connection number The first stability constant of the 1:1 complexes of the studied compounds with divalent metal ions is logK ₁ Zn(II) Cu(II) Fe(II) Fe(III) Mg(II) Ca(II) 2 4.84 4.17 5.08 5.29 2.59 2.06 3 5.40 5.32 5.95 6.59 4.36 4.57 eleven 5.05 4.39 6.27 6.60 2.82 <0.5 12 5.46 4.62 6.40 6.10 3.72 3.60

Table 5
Stepwise stability constants of complexes of 1:1 composition of the studied compounds with zinc ions, found using the Bjerrum method (aqueous solution, C _L =0.02 M, ratio M ^z+ To L 1:6, ionic strength of solution 0.5 (KNO ₃ ), 22ºС) Connection number Stepwise stability constants for complexes of the 1:1 composition of the studied compounds with zinc ions First stability constant, logK ₁ Second stability constant, logK ₂ Third stability constant, logK ₃ 2 4.84 4.50 2.60 3 5.40 5.05 3.80 6 5.62 5.15 4.04 8 6.25 5.80 5.05 10 5.50 5.24 4.48 eleven 5.05 4.70 2.87 12 5.46 5.03 3.38

Thus, the studies have shown that the tested compounds, derivatives of bisamides of dicarboxylic acids, are effective complexing agents, for most of which high values of logK ₁ ≥5 with respect to transition metal ions were found. In this case, a slightly increased complexing ability of the proposed compounds of general formula I with respect to iron ions is observed.

Claims (4)

1. Pharmaceutical composition for the prevention and/or treatment of inflammatory diseases, comprising an effective amount of a compound of formula 11 or a pharmaceutically acceptable salt thereof. 2. A medicinal product for the prevention and/or treatment of inflammatory diseases, comprising a compound of formula 11 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.