WO2008035998A1 - Medicinal composition for inhibiting propagation of human immunodeficiency virus - Google Patents

Abstract

The invention relates to medicinal compositions based on glycyrrhizic acid which makes it possible to increase the bioaccessibility of compositions and is used in medicine, virology and pharmacology, for example, in the form of an inhibitor used against the propagation of a human immunodeficiency virus. Said invention increases the bioaccessibility of a biologically active component embodied in the form of a glycyrrhizic acid derivative, for example di- and tri-nicotinate of glycyrrhizic acid. The inventive medicinal composition used for inhibiting propagation of a human immunodeficiency virus comprises sodium carbonate and glycyrrhizic acid derivatives in the form of di- and tri-nicotinate of glycyrrhizic acid of general formula I, wherein R is II or H, in which two and /or three of said R are II. The mass ratio of di- and/or tri-nikotinate of glycyrrhizic acid and sodium carbonate in a composition ranges from 3:1 to 1:3.

Description

 MEDICINAL FORMULATION FOR INHIBITING REPRODUCTION OF HUMAN IMMUNODEFICIENCY VIRUS

The invention relates to pharmaceutical compositions based on derivatives of glycyrrhizic acid, which can significantly increase the bioavailability of the compounds and can be used in medicine, virology, pharmacology, for example, as an inhibitor of the reproduction of the human immunodeficiency virus.

It is known that glycyrrhizic acid is the main ingredient in the extract of the roots of licorice (Glusurrhiza glabra) and Ural licorice (Gl. Uralepsis), has high biological activity and is a highly effective tool in the treatment of various diseases. So, it is used in the treatment of viral hepatitis, is included in the list of drugs 15 (which has about 50 items) that suppress the reproduction of human immunodeficiency virus in vitro. The named acid has an antiviral, anti-inflammatory, antipruritic and immunomodulatory effect, acts on different types of DNA and RNA viruses of the vitro and ip vivos (Varicella zoster; Nepres simrlex 1 and 2 types;

twenty . various types of human papillomavirus, including oncogenic), interrupts the replication of viruses in the early stages, causes the virion to exit the capsid, thereby preventing its penetration into the cells, which is associated with selective dose-dependent inhibition of phosphorylating kinase P.

~ r Glycyrrhizic acid interacts with the structures of the virus (possibly with proteins), changing various phases of the virus cycle, which is accompanied by irreversible inactivation of viral particles that are in a free state outside the cells, blocks the introduction of active viral particles into the cell, and violates the ability of the virus to induce synthesis

30 new viral particles, induces the formation of interferon, which is one of the components of the antiviral effect, inactivates these viruses in non-toxic to normally functioning cells concentrations. Mutant strains of viruses resistant to acyclovir and iodouridine are also highly sensitive to glycyrrhizic acid, as are non-mutant strains. The anti-inflammatory activity of glycyrrhizic acid is combined with a stimulating effect on humoral and cellular factors of immunity.

Derivatives of glycyrrhizic acid, as well as the acid itself, have high biological activity. In particular, the mono-ammonium salt of α-glycyrrhizic acid (glycyram, G) or penta-O-nicotinate of glycyrrhizic acid, 3- (b-D-2,3 ₅ 4-thi-o-icotinyl glucose pyranozyl) l 2 (a -D-3, 4-di-O-nicotinylglucypyranosyl glycyrrhetic acid (niglizine, NG) are used to inhibit the reproduction of the Marburg virus (RU, 2088232, publ. 08.27.1997).

Other glycyrrhizic acid derivatives are known that exhibit properties useful for the treatment of various diseases, which indicates the high potential of these substances for use in medicine, pharmacology, and biology.

The closest technical solution (prototype) is a derivative of glycyrrhizic acid - a glycopeptide of glycyrrhizic acid with anti-AIDS activity (RU, 2024546, publ.

12/15/1994).

One of the most important characteristics for candidates for antiviral drugs is not only the antiviral activity of the compound, but its bioavailability, i.e. the ability to be absorbed by the body. Many derivatives of glycyrrhizic acid, including the prototype compound, have poor solubility in water, and hence insufficient bioavailability when introduced into the body.

For this reason, the selection of additional components ^• to improve the bioavailability of glycyrrhizic acid derivatives during oral administration is an urgent task. The technical result of the invention is the development of such a medicinal composition that provides increased bioavailability of a biologically active component - a derivative of glycyrrhizic acid, for example, glycirrhizic acid di- and trinicotinates.

The specified technical result is achieved by the fact that in the medicinal composition for inhibiting the reproduction of the human immunodeficiency virus, including derivatives of glycyrrhizic acid, according to the invention, it additionally contains sodium carbonate, and as derivatives of glycyrrhizic acid it contains di - and / or three glycyrrhizic acid nicotinates having general formula:

R

то или H, причем два и / или три из названных R - этоwhere R - e

either H, and two and / or three of these R are

причем, массовое соотношение в составе ди - и/или три никотинатов глицирризиновой кислоты и карбоната натрия находится в интервале от 3: 1 до 1:3.

moreover, the mass ratio of di - and / or three nicotinates of glycyrrhizic acid and sodium carbonate is in the range from 3: 1 to 1: 3.

With an increase in the indicated ratio of the components more than 3: 1, the solubility of the di - and / or three nicotinates of glycyrrhizic acid is significantly reduced, and with a decrease in the ratio of the components less than 1: 3, the stability and biological activity of the drug composition decreases.

The compounds of di - and / or trinicotinates of glycyrrhizic acid are an inhibitor of the reproduction of the human immunodeficiency virus, for which the application is filed (RU, 2006108329/04, filing date 03/17/2006).

The synthesis of di - and tricinate of glycyrrhizic acid is carried out in accordance with the following scheme:

To implement this scheme, nicotinic acid is dissolved in an organic aprotic solvent by heating the reaction mixture to form a suspension, to which phosphorus pentachloride is also added while heating, then the suspension is cooled. Glycyrrhizic acid is added to the cooled suspension and then heated again. After the completion of the reaction and cooling of the obtained reaction mixture, ice and concentrated hydrochloric acid are added to it until a slightly acidic reaction. Glycyrrhizic acid di- and trinicotinates precipitate as a mixture of them. If necessary, dinicotinates can be separated from trinicotinates and are thus isolated separately. Studies of the toxicity of glycyrrhizic acid di- and trinicotinates showed that they are non-toxic and can be used in medicine, pharmacology and biology

However, glycyrrhizic acid di- and tri-nicotinates are water-insoluble and, as a result, their bioavailability is low when Heros is administered.

As mentioned above, in order to increase the bioavailability of glycyrrhizic acid di- and trinicotinates, an optimal drug composition was developed that provides a bioavailability of the compound by more than 6 times, which does not impair the anti-HIV activity of di

- and trinicotinates of glycyrrhizic acid.

As a result of studies, it was found that when using a composition containing a mixture of di - and trinicotinates of glycyrrhizic acid with sodium carbonate, the bioavailability of the target drug - di - and trinicotinates of glycyrrhizic acid significantly increases compared to the original drug without salt additives (table 1).

The addition of sodium carbonate in these ratios does not affect the stability of the drug and its biological activity. A study of the bioavailability of the claimed pharmaceutical composition is presented in two dosage forms, one of which is in the form of a suspension for administration of re оs, the other is water-soluble for intravenous administration.

The study used white Wistar male rats. Blood samples from rats were taken from the tail vein at 0, 1, 3, 5, 10 and 24, 48 and 72 hours after a single injection of the drug

To assess pharmacokinetic parameters, experimental animals received a drug substance at a dose of 7.8 mg / kg once intragastrically or intravenously. Whole blood was used as biological material to evaluate pharmacokinetic parameters.

Samples until quantification were stored at -20 ⁰ C. We used chloroform, methanol, hexane, and LiClO with the qualification of ChP and acetonitrile of the qualification “for high-performance] liquid chromatography), the Nucleosil 100-5-C18 5 microliter sorbent manufactured by MASNEREY-NAGEL (Germany), the chromatograph Milix A-02 (CJSC Milimprom A-02) Ekonova ”, Novosibirsk), beta counter 121 Raskbeta (LKB).

Example 1. The study of the stability of di- and trinicotinato glycyrrhizic acid with the addition of various amounts of sodium carbonate.

The necessary amounts of "Na ₂ CO ₃ are dissolved in a glass flask in 10 ml of water _, after which 1 mg (7x10 ^{~ 7} mol) of di- and trinicotinato-glycyrrhizic acid are added, the mixture is thoroughly mixed until dissolved. The increase in the content of Na ₂ CO _{3 is} higher than the mass ratio 1 : ^' . leads to degradation of the drug. A decrease in the content of Na ₂ CO ₃ lead to' an increase in dissolution time (table 1).

The stability of the product was monitored by HPLC: phases, LiСhroсpher 100-5-RP-18; mobile phase - 0.1% trifluoroacetic acid i acetonitrile

Example 2. A study of the bioavailability of various ratios of sodium carbonate and di- and trinicotinates of glycyrrhizic acid.

When rats are administered intragastrically to a dose of 7.8 mg / k (corresponding to a dose of 100 mg assumed for clinical trials, the maximum concentration in rat blood is reached after 8.9 hours after a single dose of administration, half-absorption period is 5.3 hours. The fraction of bioavailable dose varies in depending on the ratio of sodium carbonate and glycyrrhizic acid di- and trinicotinates i is from 12% (in the absence of sodium carbonate) to 68% (with a mass ratio of 1: 3) (table 2). Amma ASKID, two-chamber model. Table 1. The effect of the ratio of di- and trinicotinates of glycyrrhizic acid and sodium carbonate on the stability of the compound.

Table 2. The effect of the ratio of di- and trinicotinates of glycyrrhizic acid and sodium carbonate on bioavailability with intragastric administration to rats ..

Table 3. The effect of sodium carbonate on the anti-HIV activity of glycyrrhizic acid di- and trinicotinates in MT-4 cell culture.

Как видно из таблицы 3 для повышения биодоступности ди- и три- никотинатов глицирризиновой кислоты заявляемый оптимальный лекарственный состав обеспечивает повышение биодоступности соединения более чем в 6 раз. При этом данный состав не ухудшает анти-ВИЧ активности ди - и триникотинатов глицирризиновой кислоты.

As can be seen from table 3, to increase the bioavailability of glycyrrhizic acid di- and trinicotinates, the claimed optimal pharmaceutical composition provides a more than 6-fold increase in the bioavailability of the compound. However, this composition does not worsen the anti-HIV activity of di - and trinicotinates of glycyrrhizic acid.

Claims

CLAIM A medicinal composition for inhibiting the reproduction of the human immunodeficiency virus, including derivatives of glycyrrhizic acid, characterized in that it additionally contains sodium carbonate, and as derivatives of glycyrrhizic acid it contains di - and / or three glycyrrhizic acid nicotinates having the general formula: where R - e

either H, and two and / or three of these R are

moreover, the mass ratio of di - and / or three nicotinates of glycyrrhizic acid and sodium carbonate is in the range from 3: 1 to 1: 3.