RU2572800C1 - New formulation containing polyethylene glycol (peg) conjugated interferon alpha-2beta characterised by less painful administration - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine and concerns a composition for subcutaneous administration, containing PEGylated interferon alpha and additives, particularly edetate disodium dehydrate, sodium acetate trihydrate, glacial acetic acid, an osmotic agent. The group of inventions also concerns a method for producing the above composition; a kit comprising a syringe or flask filled with the above composition, and an instruction for use.

EFFECT: group of inventions provides less painful administration of the composition.

15 cl, 7 ex, 5 dwg, 13 tbl

Description

BACKGROUND OF THE INVENTION

Introduction

Interferons (IFN) are a group of biologically active proteins or glycoproteins produced by various cells in response to a viral infection or when certain chemical and biological substances are exposed to cells (Isaacs & Lindeman, 1957; Pestka et al., 2007). The binding of IFN to cellular receptors leads to the induction of the synthesis of a number of intracellular proteins that mediate the antiviral, immunomodulatory and antiproliferative effects of IFN (Pestka et al., 2004; Bekisz, 2004).

The genes of various human IFNs were cloned and expressed in bacterial and animal cells; as a result, many recombinant IFNs were obtained and purified (Pestka et al., 2004; Pestka, 2007). Medicines containing recombinant IFNs are used in clinical practice to treat a range of viral, oncological and immune diseases (Pestka et al., 2004; Chevaliez & Pawlotsky, 2009).

Currently, IFN-based drugs are used in clinical practice to treat a range of viral, oncological and immune diseases (Pestka et al., 2004). Ifn drugs were most widely used for the treatment of viral hepatitis (Chevaliez & Pawlotsky, 2009), which represent one of the most serious social problems. Currently, there are about 350 million chronically ill hepatitis B patients and 170 million hepatitis C patients worldwide (Marcellin, 2009). In most cases, hepatitis C disease acquires a chronic course with the formation of severe outcomes - liver cirrhosis and hepatocellular carcinoma (Modi & Liang, 2008). According to the WHO, since 1961, chronic hepatitis and cirrhosis of the liver, as a cause of death, have moved from 10th to 5th place in the USA and Western Europe (Marcellin, 2009). The use of preparations containing IFN-alpha is also known for the treatment of a number of leukemia and solid tumors, including recurrent melanoma (Bukowski et al., 2002; Decantris et al., 2002; Qintas-Cardama et al., 2006).

The efficacy of native IFN drugs is limited by rapid absorption from subcutaneous tissues, large distribution volume, relatively low stability, short half-life, high immunogenicity and toxicity (Wills, 1990). As a result, within a few hours after administration, a rapid decrease in the concentration of IFN in the blood plasma is observed, while interferon is not detected in the plasma 24 hours after the injection (Chatelut et al., 1999). A decrease in the concentration of interferon creates the conditions for the resumption of virus replication and an increase in the concentration of viral particles (Lam et al., 1997). Therefore, there is a need for frequent injections of the drug to achieve effective therapeutic plasma concentrations, which leads to dose-related side effects. Due to these characteristics, monotherapy of chronic hepatitis C (CHC) IFN-2b for 12 months led to a stable virological response in only 15-20% of patients; in patients with genotype Ib and high virological load, the drug was not observed at all (Mc Hutchinson et al. , 1998).

The therapeutic efficacy of IFN can be enhanced by the use of sustained release drugs, in particular “PEGylated” IFN (Manns et al., 2001; Hadziyannis et al., 2004; Zeuzem et al., 2001).

The preparation of PEGylated IFN consists in the chemical binding of an interferon molecule to a polymer - monomethoxypolyethylene glycol (mPEG), consisting of repeating subunits of ethylene oxide with a methoxy group at one end and a hydroxyl group at the other. The mPEG molecule may have a different molecular weight and stereochemical structure (linear or branched). To carry out the PEGylation reaction, the hydroxyl group at the end of the MPEG is activated by various reactive functional groups. An activated MPEG can covalently bind to a protein in one or more places, depending on the nature of the activated group and the reaction conditions (Zalipsky & Hums, 1997; Roberts et al., 2002).

PEGylation of IFN leads to an improvement in pharmacokinetics, an increase in the half-life of blood, a decrease in blood concentration fluctuations, a decrease in immunogenicity and toxicity, an increase in in vivo activity (with a decrease in in vitro activity); increased stability (Glue et al., 2000; Reddy et al., 2001).

Various PEG-IFN conjugates are known in the art.

Known conjugate PEG-IFN-2a with linear PEG with a molecular weight of 1-5000 Da (Patent US 5382657, 1995), conjugates PEG-IFN-2b obtained by conjugation of native IFN-2b with linear (patent RU 2311930, 2004) or branched (patent RU 2382048, 2008) with MPEG derivatives with a molecular weight of 13-17 kDa, and a PEG-IFN-2b conjugate obtained by binding of native IFN-2b with a branched triazine derivative of MPEG with a molecular weight of 7.5-35 kDa (Patent RU 2298560 , 2004). Also known conjugate PEG-IFN-2A, obtained by conjugation of IFN-2A with branched PEG with a molecular weight of 40 kDa (patent RU 2180595, 1997). Also known is the conjugate of PEGylated interferon-a-2b, which is a single positional isomer with monomethoxypolyethylene glycol attached to the N-terminal cysteine, with a molecular weight of PEG from 20 to 40 kDa (patent RU 2447083).

Compositions containing pegylated interferons

There are currently several types of pegylated interferon drugs on the pharmaceutical market.

So the composition of the drug "Pegasis" (manufacturer Hoffmann-La Roche, France), according to the instructions for medical use, includes 0.5 ml:

Active substance:

PEGinterferon alfa-2a (40 kDa) 135 mcg or 180 mcg

Excipients:

Sodium chloride 4.0 mg Benzyl alcohol 5.0 mg Sodium Acetate Trihydrate 1.31 mg Glacial acetic acid 0.0231 mg Polysorbate 80-0.025 mg Sodium Acetate Solution 10% up to pH 6.0 Acetic acid 10% up to pH 6.0 Water for injections up to 0.5 ml.

Another PEG-IFN-based preparation contains PegIntron (Schering-Plow, Belgium) contains:

Active substance:

PEGinterferon alfa-2beta in the form of lyophilized powder 50, 80, 100, 120 mcg

Excipients:

Sodium hydrogen phosphate, sodium dihydrogen phosphate, sucrose, polysorbate 80, water for injection.

The closest analogue of this invention can be considered a medicinal product "Algeron", having the following composition:

Active ingredient: PEGylated interferon alfa-2b (chaineginterferon alfa-2b) as an active substance  200 mg / ml

Excipients:

Sodium Acetate Trihydrate  2.617 mg / ml Glacial acetic acid  up to pH 5.0 Sodium chloride  8.0 mg / ml Polysorbate 80 0.05 mg / ml Disodium edetate trihydrate  0.056 mg / ml Water for injections  up to 1.0 ml.

The instructions for medical use report side effects expressed by the following most common symptoms: dry cough, joint pain, muscle pain, dry and peeling skin, skin itching and rash, inflammation at the injection site and others.

Preparations of interferon and its analogues or derivatives, including and PEGylated, are introduced only by injection through a syringe or a special device - an injector (syringe pen).

Often, any injection injected subcutaneously can cause reactions at the injection site in the form of soreness, redness, swelling, infiltrates, and even necrosis. The experience with the use of drugs that alter the course of multiple sclerosis (interferons beta for subcutaneous administration) indicates a high effect of tolerance at the injection site, the convenience and simplicity of injecting on long-term adherence to therapy or compliance (degree of correspondence between patient behavior and recommendations received from a doctor) . The effectiveness of interferons is dose-dependent in nature and can significantly decrease with skipped injections by the patient. Most patients report that they missed injections for various reasons: reactions at the injection site, forgetfulness, fear of needles, fear of other adverse reactions, travel. More than half of patients who refused treatment on their own, do this during the first 3.5 months from the start of therapy. The effectiveness of interferon drugs in the treatment of multiple sclerosis is limited to 30%, a key strategy to increase the effectiveness of therapy is to increase compliance.

The greatest influence of the reaction at the injection site has on:

- drugs that have a long and continuous cycle of therapy;

- drugs administered by the patient on his own on an outpatient basis;

- drugs used in pediatric practice.

Drugs with better tolerance at the injection site have advantages with long-term therapy:

- the best safety profile affects the informed choice of the drug by the patient and / or doctor (increase in the number of patients taking the drug);

- higher adherence of patients taking the drug (higher compliance), which in turn leads to the retention of a greater number of patients on therapy by reducing failures, and increasing the effectiveness of therapy due to the use of the drug without omissions, and therefore, in adequate doses;

- better tolerance is associated with a higher level of quality of life for the patient and lower costs for the treatment of patients.

In addition, during clinical trials of PEGylated interferon preparations, side effects were also reported with their administration and use. So, when using interferons and their derivatives, the most frequent observed side effects are weakness, anorexia, tremor, fever, reactions at the injection site, and depression. More severe side effects are also common, which can lead to drug discontinuation: nausea and vomiting, headache and back pain.

In addition to the described side effects, patients experience pain when administering interferon preparations. Usually this is expressed in unpleasant skin reactions: lupus-like syndrome, inflammation, pain and itching at the injection site. These sensations can persist for a long time, which, in turn, can lead to refusal of treatment by the patient. With the introduction of PEGylated interferon-alpha-2beta, eruptions in the form of erythematous vesicles at the injection site, allergic reactions and inflammations are reported.

Thus, there is a need to create a new drug containing PEG-IFN-alpha-2 beta, which would have reduced pain when administered and reduced side effects when used.

The present invention provides a pharmaceutical composition for subcutaneous administration containing PEG-IFN-alpha-2beta and excipients, which has a reduced pain when administered.

Description of figures:

FIG. 1 - A visual analogue pain scale (YOUR) is shown with a graduation in points.

FIG. 2 - VAS scores are shown with the introduction of compositions B1-B5 (averaged).

FIG. 3 - Shows YOUR points with the introduction of compositions D2 and D3.

FIG. 4 - YOUR scores are shown with the introduction of compositions A1-A5, C1-C5, D1 (averaged).

FIG. 5 - VAS scores are shown with the introduction of compositions similar in composition to the Algeron preparation and composition K.

Description of the invention

According to the present invention, a composition for subcutaneous administration, with reduced soreness, containing 1 ml:

Active substance:

PEGinterferon alpha-2b conjugate of the formula:

where m = 4 or 5, and n takes such a value that the molecular weight of PEG is 20 kDa 10-3000 mcg

Excipients:

Sodium Acetate Trihydrate 0.001-5 mg Glacial acetic acid up to pH 4.0-6.0 Osmotic agent 1-100 mg Disodium edetate dihydrate 0.001 to 0.9 mg Water for injections up to 1.0 ml.

Moreover, the osmotic agent contained in this composition is selected from the group consisting of sodium chloride, trehalose dihydrate, sucrose, sorbitol, mannitol, derivatives of mono- and disaccharides.

In addition, according to the invention, the composition has antiviral, antiproliferative and immunomodulatory activity.

According to another embodiment, the composition according to the invention can be used as a medicine for the treatment of viral and oncological diseases and diseases accompanied by primary or secondary immunodeficiency conditions. In this case, the viral disease can be hepatitis C or hepatitis B, and the cancer can be myelogenous leukemia, in particular melanoma.

Also, according to the present invention, a method for producing a composition for subcutaneous administration, including:

- preparation of a concentrate containing sodium acetate dihydrate, glacial acetic acid, osmotic agent, disodium edetate dihydrate and water for injection;

- adding conjugate PEGinterferon alpha-2b of the formula:

where m = 4 or 5,

and n takes such a value that the molecular weight of PEG is 20 kDa,

- filtering the resulting solution;

- packaging the resulting solution in dosage units.

Moreover, according to the invention, the filtration step may be a sterilizing filtration.

In yet another embodiment, a method for preparing a composition for subcutaneous administration involves packaging the resulting solution in dosage units, which may be pre-filled syringes, vials, or auto-injectors.

In another embodiment, the invention provides a kit comprising a dosage unit and instructions for use for the prevention or treatment of diseases accompanied by primary or secondary immunodeficiency states, cancer, viral diseases. Moreover, the indicated viral disease can be hepatitis C or hepatitis B, and the cancer can be myeloid leukemia, in particular melanoma.

DETAILED DESCRIPTION OF THE INVENTION

The objective of the present invention is to obtain a new composition for subcutaneous injection containing a conjugate of PEG-IFN-alpha-2beta, with antiviral, antiproliferative and immunomodulating activity (IFN-alpha activity), while this composition has reduced soreness and better tolerance when administered.

This problem is solved by selecting the qualitative and quantitative composition of the composition for subcutaneous injection, in which there is a significant reduction in pain and increased tolerance to the drug when administered.

The authors of the present invention unexpectedly found that with the introduction of a composition with a certain ratio of excipients, there is a decrease in side effects and a decrease in pain during administration. In particular, it was noted that when the content of sodium acetate trihydrate in the composition PEGinterferon alpha more than 5 mg / ml there is a sharp increase in pain with the introduction. And at the same time, when the content of sodium acetate trihydrate is less than 5 mg / ml in the composition of PEGinterferon alfa, improved tolerance to patients is observed upon administration and a decrease in pain and inflammation at the injection site.

Thus, the present invention provides a composition containing as an active substance the conjugate PEG-IFN-alpha-2beta and as auxiliary substances sodium acetate trihydrate in a concentration of not more than 5 mg / ml, glacial acetic acid, osmotic agent, disodium edetate dihydrate and water for injection in the following proportions:

Active substance:

PEGinterferon alpha-2b conjugate of the formula:

where m = 4 or 5,

and n takes such a value that the molecular weight of PEG is 20 kDa 10-3000 mcg

Excipients:

Sodium acetate trihydrate as a buffering agent 0.001-5 mg Glacial acetic acid up to pH 4.0-6.0 Osmotic agent 1-100 mg Disodium edetate dihydrate 0.001-0.9 mg Water for injections up to 1.0 ml

however, this composition has reduced soreness and better tolerance with the introduction.

As is known from previous practice for the production of injectable forms, excipients can be selected from buffering agents, osmotic agents, stabilizers, antioxidants, diluents, solvents and preservatives.

Various agents can be used as buffering agents, such as at least one substance selected from the group consisting of sodium acetate trihydrate, sodium / potassium hydro and / or dihydrogen phosphate, sodium or ammonium acetate, citrate, hydrogen carbonate, phosphate buffers.

As osmotic agents, mannitol, sodium chloride, trehalose dihydrate, sucrose, arabitol, sorbitol, glycerin, derivatives of mono- and disaccharides, lactose, dextran can be used.

As stabilizers, polysorbates, EDTA, polyvinylpyrrolidones, dextrans, HSA, para-hydroxybenzoic acid esters, alcohols (e.g. benzyl alcohol), phenols can be used.

As diluents, water and pharmaceutically acceptable alcohols may be used.

In addition, according to the present invention, the compositions can be used in the treatment of the treatment of viral and oncological diseases and diseases accompanied by primary or secondary immunodeficiency conditions. Viral diseases can be hepatitis C or hepatitis B. Oncological diseases can be myeloid leukemia, melanoma.

A method of obtaining this composition includes the steps of preparing a concentrate containing sodium acetate dihydrate, glacial acetic acid, an osmotic agent, disodium edetate dihydrate and water for injection, and further adding to it the active substance, PEGinterferon alpha-2b conjugate, filtering the resulting solution and packaging the resulting solution in dosage units. However, other methods of preparing compositions for subcutaneous administration are known from the technology. The components can be mixed in various proportions, fractionally or completely, as well as added in different sequences or simultaneously. Filtration of the solution may be a sterilizing filtration, for example, through depth or membrane filters. The pore size of the filters may be different, suitable for the sterilization process. Preferably, the pore size is selected no more than 0.22 microns. More information can be found in the injection technology manuals, in particular. Handbook of pharmaceutical manufacturing formulations.

Dosage units, according to traditional technology, can be syringes, vials or auto-injectors pre-filled with a solution of the composition according to the invention. Syringes, vials or auto-injectors can have any volume and can be made of any material suitable for this (glass, polymer material), which involves the hermetic storage of this composition for a long time.

A kit for use by a patient on an outpatient basis or in a medical facility may include a dosage unit containing a composition according to the present invention and instructions for using the composition. This kit can be used in the treatment of viral and oncological diseases and diseases accompanied by primary or secondary immunodeficiency conditions. Viral diseases can be hepatitis C or hepatitis B. Oncological diseases can be myeloid leukemia, melanoma.

The present invention is illustrated by the examples below, which are not intended to limit the invention.

Examples

Example 1

Preparation of PEG-IFN-alpha-2beta Conjugate

PEG-IFN-alpha-2 beta conjugate of the formula

where m = 4 or 5, and n takes such a value that the molecular weight of PEG is 20 kDa, obtained by the method described in patent RU 2447083.

Example 2

Obtaining compositions according to the invention for subcutaneous administration

A concentrate was prepared to obtain the final composition, including sodium acetate trihydrate, an osmotic agent, disodium edetate dihydrate, and water for injection, adjusted with glacial acetic acid to pH 4.0-6.0. Next, a PEG-IFN formula conjugate of the formula was added to the concentrate

где m=4 или 5, а n принимает такое значение, что молекулярная масса ПЭГ равна 20 кДа. Далее полученный раствор фильтровали и разливали во флаконы или шприцы. Флаконы укупоривали резиновой пробкой и обкатывали колпачками. Шприцы запаивали. Обе дозированные формы контролировали на включение механических частиц.

where m = 4 or 5, and n takes such a value that the molecular weight of the PEG is 20 kDa. Next, the resulting solution was filtered and poured into vials or syringes. The bottles were corked with a rubber stopper and dipped in caps. The syringes were sealed. Both dosage forms were controlled for the inclusion of mechanical particles.

By the method described above, compositions were obtained having the following compositions: (shown in Table 1).

Table 1. The composition of the compositions according to the invention Component Content in 1 ml Active substance No. 1-1 No. 1-2 No. 1-3 No. 1-4 No. 1-5 PEGinterferon alfa-2beta 10-100 mcg 200-500 mcg 500-1000 mcg 2000 mcg 3000 mcg Excipients: Sodium Acetate Trihydrate 0.001-5 mg 0.001-5 mg 0.001-5 mg 0.001-5 mg 0.001-5 mg Glacial acetic acid up to pH 4.0-6.0 up to pH 4.0-6.0 up to pH 4.0-6.0 up to pH 4.0-6.0 up to pH 4.0-6.0 Mannitol / Sodium chloride / Trehalose trihydrate / Sucrose / Sorbitol / Mono- and disaccharides derivatives (used one of the proposed components) 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg 1-100 mg Disodium edetate dihydrate 0.001-3 mg 0.001-3 mg 0.001-3 mg 0.001-3 mg 0.001-3 mg Water for injections up to 1.0 ml up to 1.0 ml up to 1.0 ml up to 1.0 ml up to 1.0 ml

All obtained solutions had a color from transparent to light yellow and were stable upon receipt and upon storage for 6 months.

Example 4

Packaging in a container a medicament containing a composition according to the invention

A medicine comprising an effective amount of the inventive composition obtained according to example 2 was poured in sterile conditions at 0.5, 0.8, 1.0 and 1.2 ml (for a dosage of 10-100 μg / ml), at 0.5 , 0.6, 0.75 and 0.9 ml (for a dosage of 200-500 μg / ml), 0.5, 0.6, 0.8, 1.0 and 1.2 ml (for a dosage of 500- 1000 μg / ml) in syringes made of neutral glass of hydrolytic class I, with soldered needles, covered with protective elastic or rigid caps, sealed with tips on pistons made of butyl rubber laminated with fluoropolymer. Compositions with a concentration of PEGinterferon alpha-2beta over 1000 μg / ml were poured into bottles, corked with rubber stoppers, crimped aluminum caps.

Example 5

A kit comprising a packaged drug containing the claimed composition

The kit contains 1 or 4 syringes complete with a piston (1 or 4, respectively) / bottle in a blister strip of polymer film, together with instructions for use, which are placed in a pack of cardboard.

Example 6

Compositions used to evaluate pain when administered.

All compositions were obtained according to example 2.

For the evaluation methodology, compositions with the following composition were selected:

Table 3. The composition of the compositions used to study the pain with the introduction Code of composition IN 1 IN 2 B3 AT 4 AT 5 PEGinterferon alfa-2beta 50 mcg 250 mcg 500 mcg 2000 mcg 3000 mcg Sodium Acetate Trihydrate 0.04 mg 0.1 mg 0.9 mg 1.2 mg 5 mg Glacial acetic acid Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Mannitol 1 mg 25 mg 54 mg 70 mg 100 mg Disodium edetate dihydrate 1.00 mg 1.20 mg 1.35 mg 1.50 mg 2.00 mg Water for injections Up to 1 ml Up to 1.0 ml Up to 1.0 ml Up to 1.0 ml Up to 1.0 ml

Table 4. The composition of the compositions used to study the pain with the introduction Code of composition C1 C2 C3 C4 C5 PEGinterferon alfa-2beta 50 mcg 250 mcg 500 mcg 2000 mcg 3000 mcg Sodium Acetate Trihydrate 0.04 mg 0.1 mg 0.9 mg 1.2 mg 5 mg Glacial acetic acid Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Trehalose trihydrate 1 mg 20 mg 50 mg 70 mg 100 mg Disodium edetate dihydrate 0.10 mg 0.15 mg 0.20 mg 0.30 mg 0.40 mg Water for injections Up to 1 ml Up to 1.0 ml Up to 1.0 ml Up to 1.0 ml Up to 1.0 ml

Table 5. The composition of the compositions used to study the pain with the introduction Code of composition D1 D2 D3 D4 D5 PEGinterferon alfa-2beta 50 mcg 250 mcg 500 mcg 2000 mcg 3000 mcg Sodium Acetate Trihydrate 0.04 mg 0.1 mg 0.9 mg 1.2 mg 5 mg Glacial acetic acid Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Up to pH 4.0-6.0 Sucrose 1 mg 20 mg 50 mg 70 mg 100 mg Disodium edetate dihydrate 0.10 mg 0.8 mg 0.9 mg 1.20 mg 1.45 mg Water for injections Up to 1 ml Up to 1.0 ml Up to 1.0 ml Up to 1.0 ml Up to 1.0 ml

Example 7

Assessment methodology

The study involved 30 volunteers. During the test, volunteers received subcutaneous injections in a volume of 1.0 ml into the shoulder area (one sample on the right and left). Injections were carried out in compliance with all the necessary rules of asepsis and antiseptics, by an experienced specialist using the same unmarked insulin syringes using 30G hypodermic needles. Before injection, the studied compositions had a temperature close to room temperature, the needle was inserted into the skin fold at an angle of 90 °, followed by the slow introduction of the composition.

Pain was assessed at the time of administration and 5 minutes after administration according to YOUR (visual-analogue pain scale). Any other reactions at the injection site were also recorded, including hyperemia, the formation of infiltrates, a burning sensation. The standard evaluation form is in Appendix 1. The formulations of the compositions used are presented in Appendices No. 2-5.

The visual analogue pain scale (VAS) is shown in FIG. one.

Research Results:

The study of morbidity and local irritating effects of compositions containing PEGinterferon alpha-2beta having the compositions as indicated in tables 2-5.

The study of compositions B4-B5 and D5 was completed ahead of schedule on a small number of volunteers due to the excessive soreness of these compositions.

The average value of pain at the time of administration for composition B3 was 6 points (9-3). This indicator for composition D4, B1-B2 was 5 points (7-2). Soreness 5 minutes after the introduction of these compositions had a value of 1 point (0-3). Hyperemia, edema, or other reactions at the injection site were not observed.

The study showed a high soreness of the administered formulations. Due to unsatisfactory results, other compositions were investigated.

The data are presented in FIG. 2.

According to the results of the study, it was found that compositions A1-A5, C1-C5, D1-D3 have the least pain at the injection site in comparison with other studied compositions.

Soreness at the time of administration for compositions A5 and C1-C5 was comparable and had an average of 0.44 (0-2) and 0.5 (0-2), respectively. After 5 minutes, the average pain for compositions A5 was 0.1 (0-1), C1-C5-0 (0).

When assessing the local irritant effect when applying compositions D2 and D3, mild reactions at the injection site were revealed in 4 of 9 subjects. In contrast, when administering C1-C5 compositions, one of 8 subjects showed mild itching, hyperemia of 15 minutes duration. Thus, with comparable pain indicators according to YOUR compositions C1-C5 have less local irritating effect.

For further study, you can recommend the compositions A1-A5, C1-C5 and D1-D3.

Saline solution (composition A) showed moderate soreness at the injection site, the median of pain according to YOUR at the time of administration was 2.0.

The formulations elicited mild transient reactions at the injection site. No clear differences were found for this indicator. In one subject, after administration of composition A5, discomfort at the injection site persisted for a long time.

Table 8. Soreness with the introduction of saline. n Description YOUR at the time of introduction YOUR after 5 minutes Other reactions Composition A 9 2.0 [1; 5] 0 [1; 5] - unpleasant sensation of 30 seconds Fiz. rr 2.5 (0-5) 1.0 (0-7) - fullness after 0.5 ml mild hyperemia

It should be noted that the drug for prolonged use should not have soreness that affects daily activity and interfere with daily activities. For drugs with a high injection frequency and duration of therapy, it is not advisable to use a buffer composition with soreness of more than 3 points according to YOURS.

The study involved 18 volunteers. The median pain at the time of administration for composition D2 was 4.0 [3.0; 5.0] and 0 [0; 3.0] for composition D3. The values had statistically significant differences (MW-U test, p = 0.015). The median soreness 5 minutes after administration had a value of 0 [0: 0] for each group. Indicators of pain after 5 minutes did not differ. The data are presented in FIG. 3.

The compositions elicited mild transient reactions at the injection site, see Table 9. No clear differences were found for this indicator.

According to the results of the tests, composition D2 has a significantly better tolerance in terms of pain intensity according to YOUR at the time of administration. According to the indicators of pain 5 minutes after administration and reactions at the injection site, the compositions have good tolerance and do not have significant differences between themselves.

The study of compositions A1-A4, C1-C5, D1

The study involved 30 volunteers. The summary data are presented in the table, in charts 9 and 10. Composition A1 had a minimal soreness. In most cases, the conduct of the composition was not accompanied by any sensations; the subjects specified whether the composition was administered to them or not. With the introduction of composition D2, four subjects noted intense pain at the injection site (8 YOUR points), 3 subjects noted mild soreness at the injection site (indicating 2 points). The median of pain according to YOUR for this composition was 8 points, the average value was 5.4 points. Compositions C5 and C4 had values of 2.5 [2; 3.5], 3.0 [2; 3], respectively. When conducting the Kruskell-Wallis analysis of variance when comparing the indicators of 4 groups, the values were statistically significantly different, p = 0.0001. Median test p = 0.03.

Median pain 5 minutes after administration did not differ between groups. The compositions caused mild transient reactions at the injection site, see table 10. Compositions C4 and C5 were worse tolerated.

Table 10. Soreness with the introduction of compositions A1-A5, C1-C5, D1. N Description YOUR at the time of introduction YOUR after 5 minutes Other reactions A1 10 0 [0; 0] 0 [0; 0] were absent A2-A5, C1-C3 8 2.5 [2; 3.5] 0 [0; 1.0] - light feeling of rubbing - slight discomfort at the injection site D1 7 8.0 [2; 8] 0 [0; 1.0] - unpleasant, breaking sensations in the hand - numbness of the hand - mild hyperemia 3 × 1 cm. - sharp pain at the time of administration C4, C5 5 3.0 [2; 3] 0 [0; 0] mild itching

According to the results of the studies, composition A1 has a significantly better tolerance in terms of pain intensity according to YOUR at the time of administration. According to the indicators of pain 5 minutes after administration and reactions at the injection site, the compositions have good tolerance and do not have significant differences between themselves.

However, all compositions A1-A5, C1-C5 and D1-D3 can be used with long-term treatment, because with their introduction, YOUR points do not exceed 3.

Example 8

Determination of pain with the introduction of a composition similar to the drug "Algeron"

The study involved 30 volunteers. Were investigated compositions having a composition similar to the drug "Algeron", and containing polysorbate-80. The composition is shown in Tables 11 (an alternative composition containing polysorbate-80) and Table 12 (the composition is similar to Algeron, selected as a prototype).

Table 11. The composition containing polysorbate-80. The composition of composition K Sodium Acetate Trihydrate 0.23 mg Sodium chloride 8.0 mg polysorbate 80 8 mg EDTA mg 0.05 mg Glacial acetic acid up to pH 5.0 Water for injections up to 0.5 ml Table 12. The composition is similar to the drug "Algeron", selected as a prototype. The composition of the composition similar to the drug "Algeron" / Alg Sodium Acetate Trihydrate 2.617 mg Sodium chloride 8.0 mg polysorbate 80 0.05 mg EDTA mg 0.056 mg Glacial acetic acid up to pH 5.0 Water for injections up to 0.5 ml

These formulations revealed a high pain at the time of administration, and side effects that appeared within 5 minutes after administration, and therefore the study of the morbidity of these formulations was completed ahead of schedule on a small number of volunteers.

The average value of pain at the time of administration for the Alg composition was 6 points (9-3). This indicator for composition K was 5 points (7-2). Soreness 5 minutes after administration was important for composition Alg - 1 point (0-3), for composition K - 1 point (0-3). Significant hyperimia, swelling, itching, or inflammation at the injection site was observed in 10 of 30 volunteers.

Table 13. The results of the study of pain in compositions K and Alg Composition Alg Other reactions At the time of introduction In 5 minutes one 9 0 Itching - Redness and swelling 2 7 3 - inflammation 3 5 one Redness, itching four 3 0 Redness Composition K Other reactions At the time of introduction In 5 minutes one 7 one - Unpleasant sensation (aching, discomfort) 2 7 3 - Inflammation, itching 3 four 0 - Mild itching, redness for a short time four 2 0 - Itching, redness

The results are shown schematically in FIG. 5A and 5B.

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Claims (15)

1. Composition for subcutaneous administration, with reduced soreness, containing 1 ml:
Active substance:
PEGinterferon alpha-2b conjugate of the formula:

where m = 4 or 5,
and n takes such a value that the molecular weight of PEG is 20 kDa 10-3000 mcg
Excipients:
Sodium Acetate Trihydrate 0.001-5 mg Glacial acetic acid up to pH 4.0-6.0 Osmotic agent 1-100 mg Disodium edetate dihydrate 0.001 to 0.9 mg Water for injections up to 1.0 ml 2. The composition according to p. 1, characterized in that the osmotic agent is selected from the group comprising sodium chloride, trehalose dihydrate, sucrose, sorbitol, mannitol, derivatives of mono- and disaccharides. 3. The composition according to p. 1, having antiviral, antiproliferative and immunomodulatory activity. 4. The composition according to p. 1 for use as a medicine for the treatment of viral and oncological diseases and diseases accompanied by primary or secondary immunodeficiency states. 5. The composition according to p. 4, characterized in that the viral disease is hepatitis C or hepatitis B. 6. The composition according to p. 4, characterized in that the cancer is myeloid leukemia. 7. The composition according to p. 4, characterized in that the cancer is a melanoma. 8. A method of obtaining a composition for subcutaneous administration according to claim 1, including:
- preparation of a concentrate containing sodium acetate dihydrate, glacial acetic acid, osmotic agent, disodium edetate dihydrate and water for injection;
- adding conjugate PEGinterferon alpha-2b of the formula:

where m = 4 or 5,
and n takes such a value that the molecular weight of PEG is 20 kDa;
- filtering the resulting solution;
- packaging the resulting solution in dosage units. 9. A method of obtaining a composition for subcutaneous administration according to claim 8, characterized in that said filtration step is a sterilizing filtration. 10. A method of obtaining a composition for subcutaneous administration according to claim 8, characterized in that the dosage units are pre-filled syringes, vials or auto-injectors. 11. A dosage unit containing a composition according to claim 1, in the form of a syringe or vial filled with a solution. 12. A kit comprising a dosage unit according to claim 11 and instructions for use, intended for the prevention or treatment of diseases accompanied by primary or secondary immunodeficiency states, oncological diseases, and viral diseases. 13. The kit according to p. 12, characterized in that the viral disease is hepatitis C or hepatitis B. 14. The kit according to p. 12, characterized in that the cancer is myelogenous leukemia. 15. The kit according to p. 12, characterized in that the cancer is a melanoma.

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