RU2843160C2 - Pharmaceutical composition based on a binuclear anionic nitrosyl complex and its use in a tabletted form for the treatment of cardiovascular diseases and oncological diseases - Google Patents

Abstract

FIELD: medicine; chemistry; pharmaceutics.

SUBSTANCE: invention relates to the chemical-pharmaceutical industry and medicine, in particular to the technology of producing a solid dosage stable dosage form of promising donors of nitrogen monoxide (NO), spontaneously generating NO in the absence of additional thermo-, photo- and enzymatic activation, in particular water-soluble anionic binuclear nitrosyl iron complexes of general formula Q₂[Fe₂(SR)₂(NO)₄], where SR are aromatic and aliphatic thiols of different structures. Stabilization of NO-donors is carried out due to the use of various auxiliary substances that are allowed for medical use, mainly serum albumin. Tabletted dosage form with modified NO release provides prolonged generation of NO from the structure of the complex. Rate and depth of NO release can be modulated by using different types of auxiliary substances or by varying their concentration.

EFFECT: invention provides extending the range of drugs that are NO donors with prolonged action and used for treating socially significant diseases.

2 cl, 1 tbl, 1 dwg

Description

The invention relates to anionic binuclear nitrosyl iron complexes and can be used in medical practice to create new generation drugs for the treatment of socially significant diseases.

In pharmacy and medicinal chemistry, various classes of exogenous NO donors are known. These include organic nitrates (nitroglycerin, isosorbide mononitrate), organic nitrites, sydnonimines (molsidomine), S-nitrosothiols, diazeniumdiolates (NONOates), oximes, CINODs, etc. [Granik V.G., Ryabova S.Yu., Grigoriev N.B. Exogenous donors of nitric oxide and inhibitors of its formation (chemical aspect) // Uspekhi Chem. 1997. No. 8 (66). P. 792-807]. These compounds have a number of disadvantages, such as the development of tolerance, the formation of toxic products, the need for activation to generate NO, etc., which affects clinical recommendations for their use in patients [Kaesemeyer W., Suvorava T. Nitric oxide is the cause of nitroglycerin tolerance: providing an old dog new tricks for acute heart failure // Journal of Cardiovascular Pharmacology and Therapeutics. 2022. (27)] [Arnold W. P., Longnecker D. E., Epstei R. Photodegradation of sodium nitroprusside: biologic activity and cyanide release // Anesthesiology. 1984. No. 3 (61). P. 254-260] [Titov V. Y, Petrenko Y. M. Proposed Mechanism of Nitrite-Induced Methemoglobinemia // Biochemistry (Moscow). 2005. No. 4 (70). [P. 473-483].

Work on the synthesis and study of the properties of a promising class of exogenous NO donors - nitrosyl iron complexes (NICs) with functional sulfur-containing ligands, which are structural analogues of the active centers of nitrosyl non-heme Fe-S proteins in the body, has been widely developed [Lehnert N. [et al.]. The Biologically Relevant Coordination Chemistry of Iron and Nitric Oxide: Electronic Structure and Reactivity // Chemical Reviews. 2021. No. 24 (121). P. 14682-14905.]. Unlike nitrates, which undergo enzymatic biotransformation involving sulfhydryl groups, NICs are coordination compounds that include neutral NO groups, which spontaneously quantitatively pass into solution upon dissociation. In addition, the advantages include the presence in their structure of biologically active thiol ligands, which are also released during hydrolysis, proximity to natural analogues and action at low therapeutic doses.

One of the well-known representatives of this class is NKJ with glutathione ligands. On its basis, prof. A.F. Vanin and co-authors created the powdered drug "Oxacom" [Rodnenkov O.V. [et al.]. Efficiency and safety of the new antihypertensive drug "Oxacom" - results of the 1st and 2nd phases of clinical trials // Eurasian Cardiology Journal. 2016. No. 3. P. 186-187]. The drug causes prolonged hypotension,

suppresses platelet aggregation, reduces the infarction zone in experimental myocardial infarction. However, the widespread use of this drug may be limited by the method of obtaining only in the form of a solution.

From this point of view, the advantage of the technology for synthesizing binuclear NCCIs obtained at the Federal Research Center for Problems of Chemical Physics and Medicinal Chemistry of the Russian Academy of Sciences is the possibility of isolating them in crystalline form. Such promising compounds include cationic and anionic water-soluble NCCIs of the structural “ m -S” type with aliphatic ligands of natural origin, neutral insoluble water-soluble NCCIs of the structural “ m -S” type with aromatic thiol ligands [Sanina N. A. [et al.]. Water-soluble binuclear cationic nitrosyl iron complexes with natural aliphatic thiolyls possessing cytotoxic, apoptotic and NO-donor activity // Russian Federation Patent No. 2441873], as well as neutral insoluble water-soluble NCCIs of the structural “ m -NCS” type with benzazaheterocyclic derivatives [Sanina N. A. [et al.]. Binuclear nitrosyl iron complexes with benzazaheterocyclic derivatives, the method for their preparation // Patent of the Russian Federation No. 2441872 C2]. In addition to the vasodilating effect, a number of the obtained NCCIs have cytotoxic and antibacterial activity [Sanina N.A., Serebryakova L.I., Shulzhenko V.S., Pisarenko O.I., Rudneva T.N., Aldoshin S.M. Use of a binuclear sulfur-nitrosyl iron complex of anionic type as a vasodilator drug. 2010. R. 15] [Sanina NA, Rudneva TN, Sulimenkov I. V, Konovalova NP Sashenkova TE, Aldoshin SM Antitumor activity of iron nitrosyl complexes: New donors of nitrogen monoxide // Ross. Khimicheskii Zhurnal. 2009. Vol.53. P.164-171]. Thus, the wide spectrum of pharmacological action of these NSCs allows us to draw conclusions about their potential for NO therapy of socially significant diseases.

In further development of the optimal dosage form of NCCI, it should be taken into account that they quickly decompose in aqueous solutions, and the presence of oxygen in the reaction mixture has a significant effect on the rate of this process [Pokidova O. V. [et al.]. Decomposition of the binuclear nitrosyl iron complex with thiosulfate ligands in aqueous solutions: Experimental and theoretical study // Inorganica Chimica Acta. 2020. (502). P.119369]. Generation of NO in such systems occurs immediately upon dissolution [Pokidova O. V., Psikha B. L., Emelyanova N. S., Gutsev L. G., Novikova V. O., Zagaynova E. A., Sanina N. A. Features of the decomposition of the nitrosyl iron complex with thiourea ligands under aerobic conditions: experiment, kinetic and quantum chemical modeling // Bulletin of the Academy of Sciences. Chemical Series. 2022. No. 8 (71). P. 1604-1612], which complicates their use in medical practice.

Thus, there is a need to obtain new standardized dosage forms with a given efficiency and low toxicity. The objective of the present invention is a technology for manufacturing a tablet form of NCI.

A method for stabilizing the glutathione nitrosyl complex has been proposed in the literature, which consists of freezing the pharmaceutical composition used to -10 ј -196°C for 0.1-24 h and lyophilizing it to a residual moisture content of 0.1-2.0%. This method allows

achieve a certain stability of the NKJ, however, the technology for obtaining the lyophilisate is relatively energy-intensive and time-consuming [Vanin A.F., Lozinsky V.I., Kapelko V.I., // Polymer composition for obtaining a stabilized form of the dinitrosyl iron complex and a method for obtaining the said form of the complex. Russian Federation Patent 2291880 C1].

An extremely interesting and new direction is the production of a tablet dosage form based on NCJ for further oral administration. This method of drug delivery is the most common and easily accessible. NCJ, when entering the gastrointestinal tract, effectively binds to mucin - a glycoprotein that is part of the mucus and has a cysteine-rich domain [Zolotova N. A. Structural and functional characteristics of mucins // Clinical and experimental morphology. 2014. No. 1. P. 66-72], which leads to their stabilization and is an undoubted advantage of this method of administering NCJ. In addition, it should be noted that when administered orally, NO released during the dissociation of NCJ in the stomach will have a gastroprotektorny effect. This is due to the fact that one of the protective functions of NO is to regulate mucus production and the functions of intestinal dipeptidases [Motavkin P. A. Nitric oxide in the digestive system // Pacific Medical Journal. 2004. No. 2. P. 13-17].

Thus, the inclusion of the active substance (NCI) in the solid dosage form (compressed tablet) allows to avoid preliminary dissolution, which will lead to rapid release of NO from the complex, ensures relatively long-term preservation of drugs in a solid state during storage and transportation, ease of use, allows to avoid additional manipulations and reduces the risks associated with compliance. Consequently, tablet forms based on NCI for internal use are of exceptional interest.

Tablets are characterized by portability, unpleasant organoleptic properties (taste, smell, coloring ability) are leveled by adding flavoring agents, combinations of incompatible pharmaceutical substances are easily implemented, there is a possibility of localizing and prolonging the action of the drug, and the sequential absorption of several drugs from a tablet at certain intervals can be regulated, etc. [Innovative technologies and equipment for pharmaceutical production: a textbook / N.V. Menshutina, Yu.V. Mishina, S.V. Alves. Moscow: BINOM, 2012. Vol. 1,325 p.].

The technology of tablet production involves the active use of pharmaceutically indifferent excipients to ensure the necessary characteristics of the drug.

Particular attention should be paid to the excipients used in the production of prolonged-release dosage forms [Kotsur J. M., Flisyuk E. V. Modern polymers in prolonged release tablet technology // Pharmacy Formulas. 2020. No. 1 (2). P. 36-43]. These include, for example, polyvinylpyrrolidone (PVP). The ability of PVP to bind and form complexes with

medicinal substances allows to obtain preparations with a slow release of active substances from the drug depot and to increase the duration of their action on the body. Films based on PVP containing NCJ and possessing antitumor and antiseptic activity are known in the literature [Shmatko N.Y. [et al.]. Synthesis and properties of polyvinylpyrrolidone films containing iron nitrosyl complexes as nitric oxide (NO) donors with antitumor and antiseptic activities // Russian Chemical Bulletin. 2015. No. 7 (64). P. 1616-1622].

Also, from the point of view of effective stabilization of NFI, bovine serum albumin (BSA) should be noted. Under physiological conditions, nitrosyl complexes are adsorbed on its surface, which leads to limited access of water and, consequently, a decrease in the rate of NFI decay, and to a more prolonged generation of NO groups [Pokidova O. [et al.]. Influence of hemoglobin and albumin on the NO donation effect of tetranitrosyl iron complex with thiosulfate // Nitric Oxide. 2020. (94). P. 69-72].

This is why we chose BSA and PVP as components of experimental dosage forms containing anionic NIC.

The objective of the invention is to create a new tablet dosage form based on NKJ for further oral use. This method of drug delivery is the most common and easily accessible.

The stated problem is solved by a pharmaceutical composition containing a class of NO donors that spontaneously generate NO in the absence of additional thermal, photo and enzymatic activation, and a stabilizer, including one of a peptide nature, with the following ingredient ratio (%, mass):

NO donor - 0.2-0.4, stabilizer and/or indifferent filler - 99.6-99.8.

The problem is also solved by a solid dosage form, primarily for oral use in the form of a tablet, including the said pharmaceutical composition, as well as auxiliary substances, with the following content of components (%, mass):

pharmaceutical composition according to item 1 - 0.2-0.4, polyvinyl pyrrolidone - 5 and lactose - the rest;

or a pharmaceutical composition according to item 1 -0.2-0.4, a stabilizer, including one of peptide nature -50 and lactose - the rest;

or a pharmaceutical composition according to paragraph 1-0.2-0.4 and a stabilizer, including one of peptide nature - the rest.

In addition, the problem is solved by using a pharmaceutical composition in solid tablet form for the treatment of cardiovascular and oncological diseases.

A pharmaceutical composition for obtaining a stabilized anionic tetranitrosyl binuclear iron complex of the general formula _Q2 [ _Fe2 (SR) ₂ (NO) ₄ ], where SR are aromatic and aliphatic thiolyls of various structures, in the form of a solid dosage form - pressed tablets with modified (extended) release; using albumin in various concentrations as the main auxiliary substance (prolonger).

Detailed description of the invention

The present invention relates to the production of a tablet form of tetranitrosyl anionic NICs of the composition Q ₂ [Fe ₂ (SR) ₂ (NO) ₄ ], where SR are aromatic and aliphatic thiolyls of various structures. Preferably, SR is thiosulfate.

The state of the art does not contain information on previously declared tablet dosage forms based on NKJ and the method for their production.

The method for obtaining a stabilized anionic binuclear NCJ in the form of a solid dosage form consists of grinding the solid substances included in the tablet, mixing the resulting powders to ensure uniform mixing, moistening the tablet mass (solvent - anhydrous alcohol), molding the tablet by pressing and drying under mild conditions.

Preferably, the process is carried out at room temperature, preferably at 18-25°C.

In the production of the pharmaceutical substance of water-soluble NCZ, unlike other representatives of this class of compounds, their aqueous and alcohol solutions will be used, which will avoid the use of toxic organic solvents.

Any pharmaceutically inert substances that give the appropriate form to the composition, do not weaken the therapeutic effectiveness of the NSC and are approved for medical use are used as an auxiliary substance. Preferably, serum albumin.

The pharmaceutical composition is a solid dosage form preferably for oral administration and can be presented in a set in the form of tablets, capsules in blisters, ampoules, bottles, vials, sachets, etc.

Solid dosage forms may be uncoated or may be coated using known methods, for example to delay disintegration and absorption.

The following examples of the preparation of a tablet dosage form are given only as an additional illustration of the invention and should not be considered as limiting the invention.

Examples

An anionic tetranitrosyl complex with a thiosulfate ligand of the composition Na ₂ [Fe ₂ (S ₂ O ₃ ) 2 (NO ₄ ] 4H ₂ O (complex 1) [Sanina N. A. [et al.]. Synthesis, Structure and Solid- Phase Transformations of Fe Nitrosyl Complex Na ₂ [Fe ₂ (S ₂ O ₃ ) ₂ (NO) ₄ ] 4H ₂ O // Russian Journal of Coordination Chemistry. 2005. No. 5 (31). P. 323-328], which has a vasodilating [Sanina N. A. [et al.]. Use of a binuclear sulfur-nitrosyl iron complex of anionic type as a vasodilator drug // Russian Patent No. 2437667 C1] and antimetastatic effect [Sanina N. A., Rudneva TN, Sulimenkov IV, Konovalova, NP Sashenkova TE, Aldoshin SM, Antitumor activity of iron nitrosyl complexes: New donors of nitrogen monoxide//Ross Khimicheskii Zhumal, 2009, No. 53. P. 164-171.], and is promising for use as a drug for the treatment of socially significant diseases.

Experimental solid dosage forms (tablets) of the following composition (per 100 mg, wt.%) were prepared:

1) Tablets containing 0.3% complex 1 and 5% PVP:

Complex 1 (active ingredient - active substance) - 0.3;

Polyvinylpyrrolidone (PVP) - 5.0;

Lactose - 94.7.

2) Tablets containing 0.3% complex 1 and 50% albumin:

DV-0.3;

Bovine serum albumin (BSA) - 50.0;

Lactose-49.7.

3) Tablets containing 0.3% complex 1 and 99.7% albumin:

DV-0.3;

Bovine serum albumin (BSA) - 99.7.

Tablets of this composition have not previously been described anywhere in the literature.

The iron contained in the NKJ is presented in tablets in small quantities (-0.06 mg), which do not cause side effects and toxic effects inherent in drugs available on the market and used to treat iron deficiency anemia [Qu X., Gorenkov R. V. The Toxic Effects of Ethylene Glycol Tetraacetate Acid, Ferrum Lek and Methanol on the Glutathione System: correction options // Expert Review of Clinical Pharmacology. 2021. No. 1 (14). P. 131-139].

Study of NO-donor activity of tablet form

It is known that the main product of NO oxidation in aqueous aerobic solutions is nitrite [Ignarro LJ [et al.]. Oxidation of nitric oxide in aqueous solution to nitrite but not nitrate: comparison with enzymatically formed nitric oxide from L-arginine. // Proceedings of the National Academy of Sciences. 1993. No. 17 (90). P. 8103-8107]. Figure 1 shows the kinetic dependences of NO2" accumulation in a Tris-HCl buffer solution (pH = 7.0) with a complex concentration of 6.07×10 ^-5 M. The micromolar concentration of nitrite ions in the system can be determined using UV spectroscopy according to the Griess reaction. The method is based on the diazotization of sulfanilamide with NO ₂ ion and subsequent reaction with N-(1-naphthyl)-ethylenediamine to obtain an intense pink product with an absorption maximum at λ _max = 540 nm [Moller M. N. [et al.]. Detection and quantification of nitric oxide-derived oxidants in biological systems 11 Journal of Biological Chemistry. 2019. No. 40 (294). P. 14776-14802].

The obtained dosage forms were dissolved under physiological conditions (in 0.05 M Tris-HCl buffer solution, pH=7.0, with constant stirring using a magnetic stirrer at 250 rpm and a temperature of 23°C). The amount of NO groups released during dissolution of tablets with albumin (Table 1, Fig. 1) is close to the amount of NO groups (17 mM) generated by complex 1 during its dissolution in an equimolar concentration of BSA. In contrast to the experiments with tablets, in this case the maximum concentration of NO is achieved 1.5 h after the start of dissolution (for comparison, when dissolving tablets with 99.7% albumin content, the kinetics reaches a plateau after 7 h), the effective rate constant of this process is k _eff = (9.0±0.9) × 10 ^-4 s ^-1 .

The amount of NO groups released during dissolution of tablets with PVP is close to the amount of NO- groups generated by the complex during dissolution of tablets with albumin (Table 1), but the time to reach a plateau is different - in tablets with PVP it is significantly shorter. Thus, the NIC included in the composition of tablets with albumin generate NO for a longer time compared to the NIC in tablets with PVP.

Part of the manufactured series of tablets was stored at a temperature of 4-8°C in a desiccator (desiccant - zeolite). It was found that after 3 months of storage, the samples retained their NO-donor activity.

Table 1

Characteristics of experimental tablets containing complex 1

Thus, solid dosed medicinal forms based on anionic binuclear nitrosyl iron complexes have been obtained for the first time, which generate NO in a prolonged manner and do not require preliminary dissolution before use. The invention is expected to be primarily used for the treatment of cardiovascular diseases.

Claims (4)

1. A pharmaceutical composition made in the form of a tablet, characterized in that it contains an NO donor that spontaneously generates NO in the absence of additional thermal, photo and enzymatic activation, which is an anionic tetranitrosyl complex with a thiosulfate ligand of the formula Na ₂ [Fe ₂ (S ₂ O ₃ ) ₂ (NO) ₄ ]⋅4H ₂ O, a peptide stabilizer, and/or an indifferent filler including polyvinylpyrrolidone and lactose, with a ratio of ingredients, wt.%: NO donor - 0.2-0.4; stabilizer and/or indifferent filler - 99.6-99.8. 2. Use of the pharmaceutical composition according to item 1 for the treatment of cardiovascular and oncological diseases.

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