EA041580B1 - PHARMACEUTICAL NANOSUSPENSION FOR HIV THERAPY - Google Patents

Description

The human immunodeficiency virus is a retrovirus of the lentivirus genus that causes a slowly progressive disease - HIV infection [Weiss R.A. How does HIV cause AIDS. Science 1993, 260(5112), 1273-1279. Douek D.C., Roederer M., Koup R.A. Emerging Concepts in the Immunopathogenesis of AID. Annu. Rev. Med. 2009, 60, 471-84]. The human immunodeficiency virus was independently discovered in 1983 in two laboratories: at the Pasteur Institute in France under the direction of Luc Montagnier and at the National Cancer Institute in the United States under the direction of Robert Gallo. The results of studies in which a new retrovirus was isolated from the tissues of patients with AIDS symptoms for the first time were published on May 20, 1983 in the journal Science [Barre-Sinoussi F. at al. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science 1983, 220 (4599), 868871. Gallo R. C. at al. Isolation of human T-cell leukemia virus in acquired immune deficiency syndrome (AIDS). Science 1983, 220 (4599), 865-867.]. In 2008, Luc Montagnier and Françoise Barr-Sinoussi were awarded the Nobel Prize in Physiology or Medicine for their discovery of the human immunodeficiency virus.

The virus infects cells of the immune system that have CD4 receptors on their surface: helpers, monocytes, macrophages, Langerhans cells, dendritic cells, microglial cells. As a result, the work of the immune system is inhibited and acquired immune deficiency syndrome (AIDS) develops, the patient's body loses the ability to defend itself against infections and tumors, secondary opportunistic diseases occur that are not typical for people with a normal immune status. Without medical intervention, HIV causes the death of a patient on average 9-11 years after infection (depending on the virus subtype) [https://ru.wikipedia.org/wiki/Human Immunodeficiency Virus].

According to global statistics in 2015 worldwide: 36.7 million people were living with HIV, 2.1 million people were infected with HIV, 1.1 million people died from AIDS-related diseases, 78 million people were infected with HIV since the beginning of the epidemic , of which 35 million people have died from AIDS-related illnesses since the start of the epidemic.

As of December 2015, 17 million people living with HIV had access to antiretroviral therapy, up from 15.8 million in June 2015 and 7.5 million in 2010.

In 2015, 46% of all adults living with HIV had access to treatment, up from 23% in 2010.

Since 2010, the number of new HIV infections has decreased by 6%. Globally, the number of people infected with HIV in 2015 was 2.1 million compared to 2.2 million in 2010.

Compared to a peak in 2005, AIDS-related deaths have fallen by 45% and in 2015, the number of people who died from AIDS worldwide was 1.1 million. For comparison, in 2005 this number was 2 million.

HIV can be attenuated with combination antiretroviral therapy (APT), which consists of three or more antiretroviral drugs. APT does not cure HIV infection, but controls the replication of the virus in the human body and helps to strengthen the immune system and restore its ability to fight infections. With APT, the patient's life expectancy can be extended up to 70-80 years.

In 2016, WHO released the second edition of the Guidelines for initiating antiretroviral therapy and HIV pre-exposure prophylaxis.

By mid-2016, 18.2 million people with HIV worldwide were receiving APT, meaning 46% [4350%] of all people with HIV were receiving APT.

With new WHO recommendations to treat all people with HIV and offer antiretrovirals as an additional prevention option for people at significant risk, the number of people eligible for antiretroviral therapy rises from 28 million to 36.7 million. Expanding access to treatment is one of the central targets set for 2020 to end the AIDS epidemic by 2030 [http://www.who.int/mediacentre/factsheets/fs360/ru/].

Significant progress has been made in the treatment of HIV/AIDS with APT, an effective and safe regimen for the prevention and treatment of HIV. The most advanced drug currently under registration with the Russian Ministry of Health is Elsulfavirine (Elsulfavirine, Elpida®, VM1500) for HIV/AIDS APT [(a) A. Kravchenko et al. Safety and antiviral effect of Elpida (VM-1500), a novel NNRTI (+Truvada) in treatment-naive HIV1 infected patients at 24-48 weeks therapy. HIV Drug Therapy 2016, 23-26 October 2016, P024. Glasgow, UK. (b) R.L. Murphy et al. Elsulfavirine as Compared to Efavirenz in Combination with TDF/FTC: 48-week Study. CDRI 2017, Seattle, WA with-tdfftc-at-croi-2017-300409372.html], (c) Viriom announced the interim results of Phase 3 clinical trials with Elsulfavirin, which were presented

- 1 041580 at a major HIV/AIDS conference in Moscow. https://www.viriom.com/press-releases/2016/12/13/viriom-announces-interim-results-from-phase-3-clinical-trials-with-elpida-were-presentedat-largest-events- on-hivaids-in-moscow.].

Elsulfavirin/Elpida/VM-1500 (1a) is a prodrug of the active compound VM-1500A (1b), which is a potent inhibitor of HIV-1 HXB2 replication in MT-4 cells and belongs to the class of non-nucleoside reverse transcriptase inhibitors (Non-Nucleoside Reverse). Transcriptase Inhibitors - NNRTIs). The mean IC ₅₀ value obtained for VM-1500A (1b) on wild-type HXB2 strain and for inhibition of replication of mutant HIV-1 viruses containing mutations V106A, G190A, L100I/K103N and K103N/Y181C are significant for: HXB2 1.3 ± 0.4 nM, V106A 1.2 ± 0.2 nM,

G190A 0.6 ± 0.6 nM, L100I/K103N 1.3 ± 0.3 nM, K103N/Y181C 1.3 ± 0.4 nM.

where R is C ₂ H ₅ CON ^- Na ⁺ , NH2;

Despite the successes achieved in the treatment of HIV/AIDS with the help of APT, the development of more effective and safe schemes for the prevention and treatment of HIV remains relevant.

In particular, to date, there are no long-acting (LongActing - LA) drugs for long-term maintenance therapy of HIV / AIDS in the world.

This invention relates to a pharmaceutical composition (nanosuspension) for a long-acting injectable (LAI - long-acting injectable) drug for long-term maintenance therapy of HIV/AIDS.

The following are definitions of various terms used to describe this invention. These definitions apply to the terms as used in this specification and claims, unless otherwise limited in specific cases, either individually or as part of a larger group.

The term active ingredient (drug) refers to a physiologically active substance of synthetic or other (biotechnological, plant, animal, bactericidal, etc.) origin with pharmacological activity, which is the active ingredient of a pharmaceutical composition.

The term crystalline form means the structure of a substance, characterized by the packing of the molecules that form it into one of the types of a crystal lattice.

The term polycrystalline form means the structure of a substance having a polycrystalline structure, i.e. consisting of many small single crystals, i.e. crystallites of a certain crystalline form.

The term medicinal product means a substance (or a mixture of substances in the form of a pharmaceutical composition) in the form of tablets, capsules, injections, ointments and other finished forms, intended to restore, correct or change physiological functions in humans and animals, as well as for the treatment and prevention of diseases , diagnostics, anesthesia, contraception, cosmetology and other.

The term pharmaceutical composition means a composition comprising a compound of general formula 1 and at least one of the components selected from the group consisting of pharmaceutically acceptable and pharmacologically compatible excipients, solvents, diluents, carriers, auxiliary, distributing and receiving agents, means such as preservatives, stabilizers, fillers, grinders, humectants, emulsifiers, suspending agents, thickeners, sweeteners, flavors, flavors, antibacterial agents, fungicides, lubricants, sustained delivery regulators, the choice and ratio of which depends on the nature and method of administration and dosage. Examples of suspending agents are ethoxylated isostearyl alcohol, polyoxyethylene, sorbitol and sorbitol ether, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, as well as mixtures of these substances. Protection against the action of microorganisms can be provided with a variety of antibacterial and antifungal agents, such as parabens, chlorobutanol, sorbic acid, and the like. The composition may also include isotonic agents such as sugars, sodium chloride and the like. Prolonged action of the composition can be provided with agents that slow down the absorption of the active principle, for example, aluminum monostearate and gelatin. Examples of suitable carriers, solvents, diluents and delivery vehicles are water, ethanol, polyalcohols and mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters (such as ethyl oleate). Examples of fillers are lactose, milk sugar, sodium citrate, calcium carbonate, calcium phosphate and the like. Examples of grinders and spreaders are starch, alginic acid and its salts, silicates. Examples of lubricants are magnesium stearate, sodium lauryl sulfate, talc, and high molecular weight polyethylene glycol. A pharmaceutical composition for oral, sublingual, transdermal, intramuscular, intravenous, subcutaneous, topical or rectal administration of the active principle, alone or in combination with another active principle, can be administered to animals and humans in standard form of administration, as a mixture with conventional pharmaceutical carriers. Suitable unit administration forms include oral forms such as tablets, gelatin capsules, pills, powders, granules, chewing gums and oral solutions or suspensions, sublingual and buccal administration forms, aerosols, implants, topical, transdermal, subcutaneous, intramuscular, intravenous, intranasal or intraocular administration forms and rectal administration forms.

The term excipient as used herein refers to a compound that is used to prepare a pharmaceutical composition and is generally safe, non-toxic, neither biologically nor otherwise undesirable, and includes excipients that are acceptable for use in veterinary, as well as pharmacologically acceptable for human use. The compounds of this invention may be administered alone, but generally they will be administered in admixture with one or more pharmaceutically acceptable excipients, diluents or carriers, selected in consideration of the intended route of administration and standard pharmaceutical practice.

The term therapeutically effective amount as used herein means the amount of a substance, prodrug or drug required to reduce the symptoms of a disease in a subject. The dose of the substance, prodrug or drug will be tailored to individual requirements in each particular case. This dosage may vary widely depending on numerous factors such as the severity of the disease being treated, the age and general health of the patient, other drugs the patient is being treated with, the mode and form of administration, and the experience of the attending physician. For oral administration, the daily dose is from about 0.01 to 10 g, including all values in between, per day in monotherapy and/or in combination therapy. The preferred daily dose is from about 0.1 to 7 g per day. Typically, treatment is started with a large initial loading dose to rapidly reduce or eliminate the virus, followed by a tapering dose to a level sufficient to prevent a resurgence of infection.

The term subject means a mammal, preferably the subject is a human. It is assumed that any of the prodrugs of general formula 1, its isotopically enriched analogs, its pharmaceutically acceptable salts, hydrates, solvates, crystalline and polymorphic forms, or in combination with other compounds and drugs for combination antiretroviral therapy for HIV can be in the method of treating a subject. /AIDS.

Disclosure of invention

Surprisingly, the inventors have found that a pharmaceutical nanosuspension containing a compound of general formula 1 in crystalline or polycrystalline form as an active substance is effective as an injectable preparation for long-term maintenance therapy of HIV infection.

The subject of this invention is a nanocomposition comprising, as an active substance, a compound of general formula 1 in a crystalline or polycrystalline form in a therapeutically effective amount, optionally in combination with a pharmaceutically acceptable excipient, additive or diluent for the production of a pharmaceutical nanosuspension.

More preferred is a lyophilized nanocomposition comprising as active substance a compound of formula 1b in crystalline or polycrystalline form, in combination with a solubilizer (eg poloxamer P338) and a detergent (eg mannitol or sucrose).

More preferred is a lyophilized nanocomposition with a particle size of 200 to 900 nm.

The subject of this invention is also a pharmaceutical nanosuspension comprising the nanocomposition described above, phosphate buffered saline (PBS) and water for injection for long-term maintenance therapy of subjects infected with HIV/AIDS.

The subject of this invention is also a method for obtaining a lyophilized nanocomposition, which consists in rotational grinding with zirconium sand in an aqueous solution

- 3 041580 poloxamer of the compound of general formula 1 in crystalline or polycrystalline form, solubilizer and detergent, subsequent separation of zirconium sand and lyophilization of the resulting suspension.

The subject of this invention is also a method for preparing a pharmaceutical nanosuspension by mixing the lyophilized nanocomposition described above and water for injection.

This invention is illustrated by drawings.

Fig. 1 is a plot of particle size measurements of three samples after 24 hours of milling with sucrose.

Fig. 2 is a plot of particle size measurements of three samples after 24 hours of mannitol milling.

Fig. 3 is a plot of particle size measurements of three samples after 24 hours of grinding and 20 hours of settling (sucrose).

Fig. 4 is a plot of particle size measurements of three samples after 24 hours of grinding and 20 hours of settling (mannitol).

Fig. 5 is a plot of particle distribution measurements of three samples of Lot No. 740-1-101.1 (sucrose) after lyophilization and resuspension.

Fig. 6 is a plot of particle distribution measurements of three samples of Lot No. 740-1-101.2 (mannitol) after lyophilization and resuspension.

Fig. 7 - Pharmacokinetic curves of VM-1500A in dog plasma after single SC and IM administration of pharmaceutical nanosuspensions VM-1500-LAI and VM-1500A-LAI at a dose of 10 mg/kg.

The best embodiment of the invention

The present invention will now be described in connection with certain embodiments, which are not intended to limit its scope. On the contrary, the present invention embraces all alternatives, modifications and equivalents that may be included within the scope of the claims. Thus, the following examples, which include specific variations, illustrate but do not limit the present invention.

Example 1. Method for obtaining a lyophilized nanocomposition.

A Jar is charged with zirconium sand (150 ml) (grinding media: 0.5 mm YTZ® Zirconia Grinding and Dispersion Media; Tosoh USA, Inc.) and a solution of poloxamer P338 (10.0 g) and sucrose or mannitol (11.6 d) in 400 ml of phosphate buffer solution (PBS, pH = 7.4). 150 ml of the resulting solution was placed in a vessel (Jar), zirconium sand (150 ml) (grinding media: 0.5 mm YTZ® Zirconia Grinding and Dispersion Media; Tosoh USA, Inc.) and compound VM1500A (15.07 g) were added. The jar is placed on a milling apparatus (U.S. Stoneware 700 Series jar mill) and set to 104 rpm and ground for 24 hours. The end of the process is monitored by particle distribution analyzes on a Malvern Zetasizer Nano ZS instrument. At the end of the grinding process, the rotation is stopped and left for 16-20 hours at a temperature of 2-8°C to precipitate zirconium sand. The suspension is filtered through a glass filter with a porosity of 5-15 microns and analyzed to determine the content (concentration) of the VM1500A compound (92-97% of nominal). A nanosuspension is obtained, which, under sterile conditions, is poured into 2 ml glass sterile vials with a capacity of 5 ml, frozen and lyophilized. Get lyophilized nanocomposition, the characteristics of which are presented in table. 1 and 2 and in Fig. 1, 2, 3 and 4.

Table 1. Particle sizes in the nanosuspension after 24 h of grinding (values averaged over three measurements)

Sample Sample volume, µl Particle size, nm PDI* Quality Lot No. 740-1-101.1 (sucrose) thirty 442.3 0.428 good Lot No. 740-1-101.1 (sucrose) 10 357.7 0.283 good Lot No. 740-1-101.2 (mannitol) thirty 395.6 0.31 good Lot No. 740-1-101.2 (mannitol) 10 552.4 0.487 good

*PDI - Hereinafter - dispersion index characterizing the homogeneity of particles in the mixture.

Table 2. Particle sizes in the nanosuspension after 24 h of grinding and 20 h of settling (values averaged over three measurements)

Sample Sample volume, µl Particle size, nm PDI Quality Lot No. 740-1-101.1 (sucrose) thirty 402.9 0.367 good Lot No. 740-1-101.1 (sucrose) 10 362.1 0.268 good Lot No. 740-1-101.2 (mannitol) thirty 403.5 0.28 good Lot No. 740-1-101.2 (mannitol) 10 381.3 0.286 good

Example 2. Method for obtaining a pharmaceutical nanosuspension

To the nanosuspension obtained in example 1, a previously prepared sterile PBS solution with pH = 6.8 is added at the rate of 2.2 ml per 5 ml vial. Get pharmaceutical nanosuspension, ready for use, the characteristics of which are presented in table. 3 and in FIG. 5 and 6.

- 4 041580

Table 3. Particle distributions in pharmaceutical nanosuspension

Pharmaceutical nanosuspension Lot No. 740-1-101.1 (sucrose) Lot No. 740-1-101.2 (mannitol) pH 7.44 7.29 Particle distribution Particle size, nm 381.7 447.6 (10 µl sample) PDI 0.255 0.248 Particle distribution Particle size, nm 386.1 442.6 (30 µl sample) PDI 0.252 0.252 HPLC analysis (% of nominal amount) 93.5% 95.0%

Example 3 Pharmacokinetics of Pharmaceutical Nanosuspensions VM1500LAI and VM1500A-LAI in Dogs

Nanosuspensions VM1500LAI and VM1500A-LAI, obtained in example 2, were administered to Beagle dogs once subcutaneously and intramuscularly at a dose of 10 mg/kg, and the content of VM1500 in the blood plasma of dogs was analyzed for 120 days after injection. The determination of the concentration of the studied compounds was carried out using the HPLC-MS/MS method.

For dose administration, prepared nanosuspensions are administered to dogs subcutaneously or intramuscularly in a volume of 0.1 ml/kg in the same place. Blood was collected after 0, 0.5, 1, 2, 4, 8, 24 h and 2, 3, 6, 8, 12, 15, 18, 22, 29, 36, 43, 50, 57, 64, 71, 78, 85, 92, 107, 114 and 121 days after injection. Whole blood for plasma was collected in tubes containing sodium heparin. Immediately after blood sampling, the tube was carefully inverted 8 times for better mixing of blood and sodium heparin, after which it was incubated at room temperature for 15 min. Then the plasma was separated by centrifugation for 10 min at 3000 rpm. After centrifugation, a 1 ml aliquot of blood plasma was carefully transferred into a cryovial using an automatic pipette, avoiding touching the cell mass/separating gel with the spout. From one Vacuette tube, plasma samples were transferred to two cryovials. The obtained samples were frozen. Prior to analysis, plasma and FEC samples are stored at -80°C.

HPLC-MS/MS analysis was used to determine the concentration of VM-1500 and VM-1500A. The structurally similar compound VM-6819 was used as an internal standard (IS). When scanning in the total ion current (MS1) mode, the molecular ion of the studied compounds and IS were determined, the main product ions were recorded in the MS2 mode. For quantitative analysis, the MS/MS method was optimized in MRM mode to achieve maximum sensitivity. At least two MRM transitions for each analyte were recorded in the analysis, and one of them was used in the quantitative processing of chromatograms.

Chromatographic conditions were adjusted by introducing a solution of the test compound and IS in MeCN:H ₂ O (1:1) into the MS/MS detector via the HPLC system to achieve optimal chromatographic parameters. Standard calibration samples VM1500 and VM1500A in blood plasma were prepared in the concentration range 1.0-2000.0 ng/ml. In addition to calibration standards, a zero sample k0, k0 with IS and two series of control samples (QC) with low, medium, and high concentrations were prepared, which served as an indicator of the quality of quantitative calculations. From stock solutions of VM1500 (2.0 mg/ml) and VM1500A (2.0 mg/ml) in DMSO, 10-fold working solutions in MeCN:H ₂ O (1:1) were prepared. Solutions for calibration standards (k) and QC samples were prepared from separate aliquots of VM1500 and VM1500A stock solutions.

Experimental plasma samples used to prepare standard samples were thawed at room temperature, mixed and centrifuged for 5 min at 14,000 rpm. 90 µl of intact plasma was transferred into a 1.5 ml test tube, 10 µl of a 10-fold standard solution of the mixture of test substances in a mixture of acetonitrile:water 1:1 was added. To 90 µl of the experimental sample was added 10 µl of a pure mixture of acetonitrile:water 1:1. The samples were thoroughly mixed on a vortex for 10 s. 10 µl of IS (VM-6819 5 mg/ml in acetonitrile:water 1:1) was added, vortexed for 10 sec. 1 ml of a mixture of ethyl acetate:hexane (60:40) was added to the samples and mixed on a vortex for 20 s, and then on a shaker at 1000 rpm for 5 min. The samples were then centrifuged at 14,000 rpm for 10 minutes.

0.8 ml of the organic phase was taken into 1.5 ml test tubes and evaporated under nitrogen flow in an evaporator at a temperature of 40°C. Dry extracts were diluted in 150 μl of acetonitrile:water (1:1), vortexed for 10 s, and transferred to a clean plate for HPLC/MS/MS analysis.

The concentrations of VM1500 and VM1500A in the samples were calculated from the calibration curve constructed from the areas of the chromatographic peaks of the standard samples VM1500 and VM1500A in the matrix (blood plasma or FEC), normalized to the area IS. The construction of the calibration curve was carried out by selecting the simplest model that adequately describes the dependence of the analytical signal on the concentration, and using a suitable normalization. The main pharmacokinetic parameters were calculated by an out-of-model method using the computer program WinNonlin Professional 6.3 (Pharsight

- 5 041580

Corporation) based on experimentally obtained concentration-time data for each animal:

C _max - the maximum concentration of the substance in the blood plasma;

T _max - the time to reach the maximum concentration in blood plasma;

AUC ₀ . _t is the area under the pharmacokinetic curve from the moment of drug administration to the last observation point with a measured concentration;

AUC ₀ . _inf is the area under the pharmacokinetic curve from the moment of drug administration to infinity;

T _1/2 - the half-life of blood plasma;

kel - elimination rate constant - a parameter characterizing the rate of excretion of a substance from blood plasma;

MRT is the average retention time of a drug in the body from the moment the drug is taken.

Statistical processing of the results was carried out using descriptive statistics. The arithmetic mean values of the parameters (M), standard deviation (SD), standard error of the mean (SE), coefficient of variation (CV), and median were calculated. Statistical analysis was performed using WinNonlin Professional 6.3 (Pharsight Corporation).

In table. 4 summarizes the pharmacokinetic parameters characterizing the behavior of VM-1500 and VM-1500A following subcutaneous (SC) and intramuscular IM administration of VM-1500-LAI and VM-1500A-LAI nanosuspensions to dogs.

Table 4. Pharmacokinetic parameters of VM-1500 and VM-1500A in dog blood plasma after SC and IM administration of VM-1500-LAI and VM-1500A-LAI nanosuspensions at a dose of 10 mg/kg

Options Unit rev. SC IM SC IM VM1500-LAI VM1500LAI VM1500A- LAI VM1500LAI VM1500A -LAI VM-1500 VM-1500A AUC _inf h* ng/ml 4 435.7 5 743.6 89 210.0 89 409 121 175.1 83 916 AUCi _as t h* ng/ml 3 989.9 5 596.7 88 830.4 78 492 120 975.5 79 518 _Cmax ng/ml 111.4 1 158.7 558.7 104 1 225.0 153 Tsh h 229 74.9 170 581 153 446 Kei 1/h 0.003 0.009 0.004 0.001 0.005 0.002 MRT _inf h 180 38.6 205 957 90 657 MRTjust h 95 29.1 200 671 88 537 vain H 1.7 1.0 64 216 32 64.0

With intramuscular injection, higher _Cmax values of both VM-1500 itself and its metabolite VM-1500A are achieved than with subcutaneous administration. However, when using the VM-1500A-LAI formulation, this difference is not so significant. At the same time, the total exposure of VM-1500 and VM-1500A when administered intramuscularly in the form of VM-1500-LAI is higher than when administered subcutaneously, while the exposure of VM1500A does not depend on the route of administration of the VM-1500A-LAI formulation. Also, when administered intramuscularly, VM-1500A reaches its maximum concentration earlier than when administered subcutaneously. The half-life does not depend significantly on the route of administration.

Mean plasma concentrations (C) of VM-1500A in dogs after a single SC and IM administration to dogs in the form of VM1500-LAI and VM1500A-LAI nanosuspensions are shown in Table 1. 5 and in FIG. 7.

- 6 041580

Table 5. Plasma concentration of VM-1500A in dogs after a single SC and IM administration in the form of depot formulations of VM-1500-LAI and VM-1500A-LAI at a dose of 10 mg/kg

Time VM1500-LAI IM VM1500-LAI SC VM1500A-LAI IM VM1500A-LAI SC h DAYS Mean, ng/ml SD, ng/ml Mean, ng/ml SD, ng/ml Mean, ng/ml SD, ng/ml Mean, ng/ml SD, ng/ml 0 0.0 0.00 0.00 0.00 0.00 0.00 0.00 1 0.0 55.67 7.76 3.54 0.58 6.35 1.59 3.40 3.48 1 0.0 129.67 12.74 5.62 1.56 11.96 6.64 4.24 2.56 2 0.1 350.67 74.54 16.57 2.75 28.80 9.30 7.12 4.59 4 0.2 482.33 87.93 31.43 6.11 40.23 3.82 12.25 5.03 8 0.3 892.67 108.49 77.93 8.47 84.43 25.20 16.43 3.98 24 1.0 1166.67 151.85 314.67 35.64 124.50 11.46 49.80 13.17 48 2.0 1023.00 187.20 480.00 155.79 131.67 4.25 70.23 16.42 72 3.0 812.67 28.36 535.33 210.24 151.33 18.06 91.73 5.61 144 6.0 245.33 51.60 232.67 8.14 99.37 22.09 59.96 4.82 192 8.0 105.70 26.49 129.00 16.70 115.50 12.26 86.97 3.57 288 12.0 39.43 9.03 90.27 10.83 101.22 8.75 96.22 24.58 360 15.0 12.97 4.16 48.40 7.41 79.38 22.07 57.62 4.52 432 18.0 7.06 2.98 36.10 5.20 61.52 7.52 71.20 4.98 528 22.0 6.04 1.89 34.30 3.70 48.85 7.19 56.42 9.05 696 29.0 2.61 0.84 16.80 4.11 37.07 3.26 65.5 10.8 864 36.0 1.03 0.10 8.57 2.31 27.80 7.89 37.8 3.1 1032 43.0 0.89 0.18 4.83 1.98 32.37 5.09 34.5 12.5 1200 50.0 BLQ 1.47 0.70 13.30 1.31 18.4 3.9 1368 57.0 BLQ BLQ 7.6 2.55 12.02 2.48 1536 64.0 BLQ BLQ 12.27 3.52 12.40 1.60 1704 71.0 BLQ BLQ 8.88 2.74 8.53 1.12 1872 78.0 BLQ BLQ 6.03 1.84 11.24 3.41 2040 85.0 BLQ BLQ 5.87 2.07 9.92 2.52

Table 5 (continued). The concentration of VM-1500A in the blood plasma of dogs after a single SC and IM administration in the form of depot formulations VM-1500-LAI and VM-1500A-LAI at a dose of 10 mg/kg

2208 92.0 BLQ BLQ 6.81 1.46 12.79 3.01 2568 107 BLQ BLQ 6.43 1.38 9.59 0.39 2736 114 BLQ BLQ 9.68 1.33 5.72 1.12 2904 121 BLQ BLQ 5.70 1.81 11.47 1.07

BLQ - below the limit of quantitation

As can be seen from Table. 5 and FIG. 7, upon IM administration of a pharmaceutical nanosuspension of VM-1500-LAI at a dose of 10 mg/kg to dogs, the effective concentration of the drug VM1500A ( _IC50 = 1.3 nM or 0.74 ng/ml) in plasma is maintained for a month (after 29 days C _{VM -1500A} = 2.61 ng / ml), and with SC administration - within one and a half months (after 43 days C _VM-1500A = 4.83 ng / ml).

An even more convincing result is observed with IM and SC administration of VM-1500LAI nanosuspension at a dose of 10 mg/kg to dogs. In this case, the effective concentration of VM1500A is maintained for four months (after 121 days C _VM-1500A = 5.70 ng/ml with IM administration and C _VM-1500A = 11.47 ng/ml with SC administration).

Claims (4)

1. Pharmaceutical composition for injectable long-term maintenance therapy of HIV infection, including 2-[4-bromo-3-(3-chloro-5-cyanophenoxy)-2fluorophenyl]-N-(2-chloro-4-sulfamoylphenyl)acetamide as an active ingredient formula 1b with particle sizes from 200 to 900 nm lb (VM1500A, RO4970335) 2. Composition according to claim 1, comprising a solubilizer - poloxamer P338 and a detergent selected from mannitol and/or sucrose, phosphate-buffered saline (PBS) and water for injection. 3. A method for preparing a composition according to claim 1 or 2, including the step of grinding with zirconium sand in an aqueous solution of a poloxamer solubilizer, a compound of formula 1b specified in - 7 041580 1, phosphate buffer solution and detergent, followed by separation of zirconium sand to obtain a suspension, which, if necessary, is subjected to lyophilization, and the subsequent stage of mixing the resulting composition with phosphate buffered saline (PBS) with pH = 6.8 and water for injection. 4. The use of a pharmaceutical composition according to any one of claims 1 or 2 for intramuscular or subcutaneous long-term maintenance therapy of HIV infection.