RU2488383C1 - Solid dosage pharmaceutical composition having ability to reduce exudation, providing faster wound cleansing from necrotic mass, as well as providing faster epithelisation and regeneration - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to a pharmaceutical composition having an ability to reduce exudation, to provide faster wound cleansing from necrotic mass, as well as providing faster epithelisation and regeneration. The composition contains acexamic acid and target additives represented by aminoacetic acid, sorbitol, sodium cyclamate, the edible flavour Orange 9374240. The pharmaceutical composition is presented in the form of sachets.

EFFECT: new pharmaceutical composition is presented.

2 cl, 4 tbl

Description

The invention relates to medicine, in particular, to a pharmaceutical composition having the ability to reduce exudation, accelerate the cleansing of a wound from necrotic masses, and accelerate epithelization and regeneration containing Acemine - Acexamic acid, ε-acetylaminocaproic acid (hereinafter AK).

One of the current trends in modern traumatology and orthopedics is the search for new ways to optimize the reparative regeneration of bone tissue. The solution to this problem is inextricably linked with research on the morphology of the consolidation process. The successes achieved in this direction made it possible to discover a number of patterns of bone tissue repair, on the basis of which the doctrine of the staging and zonality of reparative osteogenesis was created (“Aesthetic and reconstructive surgery of the lower extremities”, A.A. Artemiev and others, under the editorship of A.A. .Artemyeva. Published in 2008, 248 pp. "Traumatology", under the editorship of GP Kotelnikov, SP Mironov. 2008, 808 pp. "Traumatology", under the editorship of GP Kotelnikov , S.P. Mironova, V.M. Miroshnichenko. 2006, 400 p.).

Despite the fact that the authors, using all kinds of criteria and approaches, describe the various stages of this process, the common thing for everyone is the isolation of two main phases - the phase of formation of connective tissue callus and its subsequent mineralization. This division is purely conditional, since bone regeneration is a continuous transition of one stage to another, with the prevalence of certain metabolic and morphological processes at the moment.

The accumulated experimental data allow us to conclude that all elements of bone tissue participate in osteogenesis: periosteum, havers canals, endosteum, bone marrow and surrounding tissues.

According to the cellular composition, bone regeneration sources are: fibrocytes, chondrocytes and chondroblasts, osteocytes and osteoblasts, histiocytes, lymphoid cells, fat cells, elements of myeloid and erythropoiesis, reticulocytes and endothelial cells.

Most researchers (“Traumatology”, under the editorship of G.P. Kotelnikov, S.P. Mironov. 2008, 808 pp. “Traumatology”, under the editorship of G.P. Kotelnikov, S.P. Mironov, V. M. Miroshnichenko. 2006, 400 pp. Khusid I.Kh. “The role of thyrocalcitonin in reparative osteogenesis”: Diss. Candidate of medical sciences. - Kalinin, 1975.) divide bone restoration into four stages:

Stage 1 - catabolism of tissue structures, dedifferentiation and proliferation of cellular elements;

Stage 2 - progressive proliferation and differentiation of cellular elements that secrete the organic basis of the bone matrix;

Stage 3 - complete restoration and intensification of the vascular supply of the emerging bone regenerate and mineralization of the protein base of the regenerate, which together provide the appearance of the primary bone structure;

Stage 4 - restoration of the original structure and function of the bone, physiological homeostasis.

The first three stages are carried out rather quickly, ensuring restoration of the anatomical integrity of the bone organ, and the last proceeds at a slower pace, as it determines the functional reconstruction of bone tissue.

Currently, AK is used to treat long-term non-healing wounds, burns, as well as for teary fractures, especially with prolonged tissue non-growth, to accelerate the formation of a postoperative cosmetic scar.

Applied externally (in the form of a 5% ointment or lotion), as well as inside (in the form of a 25% solution) (M.D. Mashkovsky, Medicines, M., New Wave LLC, 2002, v.2 p. 169).

In addition, it is known to use AK in combination with other drugs (lidocaine, ofloxacin) in the form of a spray for the treatment of wound surfaces (RF patent No. 2416394). Known pharmaceutical composition for local use, with antibacterial and necrolytic effects, containing as one of the components of AK, made in the form of a soft dosage form (ointment, or gel, or suppository, or capsule for rectal administration (RF patent No. 2367469).

A number of authors ("Synergistic effect of beta-cryptoxanthin and zinc sulfate on the bone component in rat femoral tissues in vitro: the unique anabolic effect with zinc", Biol. Pharm. Bull., 2005, Nov; 28 (11), p. 2142-5. "Increase in bone growth factors with healing rat fractures: the enhancing effect of zinc", Int. J Mol Med., 2001, Oct; 8 (4), p.433-8. "Effect of essential trace metal on bone metabolism in the femoral-metaphyseal tissues of rats with skeletal unloading: comparison with zinc-chelating dipeptide ", Calcif. Tissue Int., 1996, Jul; 59 (1), p. 27-32." Stimulatory effect of beta- alanyl-L-histidinato zinc on alkaline phosphatase activity in bone tissues from elderly rats: comparison with zinc sulfate action ", Biol Pharm Bull., 1994, 17 (2), p.345-7." Reviewbeta-Alanyl-L-histidinato zinc and bone resorption ”, Gen Pharmacol, 1995, Oct) indicate in their works the possibility of using AK as stimulants recovery process of bone damage. However, a more detailed study of the mechanism of the effect of AK on the various stages of healing of fractures of the tubular bones is necessary.

The closest in technical essence is the analog - Acexamic acid: instructions, use and formula. Radar Reference (http://www.rlsnet.ru/mnn_index_id_1901.htm).

Its disadvantage is that the powder is not in dosage form.

Therefore, the search and development of a new original pharmaceutical composition combining a single dosage with convenient packaging is very relevant.

The goal is achieved by a solid dosage pharmaceutical composition containing AK, excipients, with the ability to reduce exudation, accelerate the cleaning of the wound from necrotic masses, and also accelerate the epithelization and tissue regeneration, made in the form of sachets.

AK powder for sachets is the original dosage form.

The development of the composition of the dosage form - powder consisted of several stages:

- the study of the physicochemical and structural-mechanical properties of the substance;

- development of the composition of the dosage form, taking into account the lack of chemical interaction between the components and the technology for the preparation of a homogeneous powder;

- production of experimental batches of the preparation AK powder for analytical, pharmacological and toxicological studies.

Taking into account the studied technological properties of the substance, a selection of excipients was carried out.

Sorbitol was chosen as the main auxiliary substance. Aminoacetic acid was added to improve the flowability of the powder. Sodium cyclamate and orange flavor were used as flavor correlators.

Based on experimental studies of the quality of the obtained powder, its optimal composition is established in the following ratio of components, wt.%:

Acexamic acid (AK) 45.0-55.0 Aminoacetic acid 1.80-2.20 Sorbitol 39.24-47.96 Sodium Cyclamate 3.60-4.40 Fragrance food Orange 9374240 0.36-0.44

The technology for producing this dosage form is simple: all components are sifted out from possible mechanical impurities, the powder in an amount of 5 g ± 3.0% of the total mass is placed in bags from the combined Buflen material or from packaging paper with a polyethylene coating.

The following examples illustrate the invention.

Table 1 Components, g Examples one 2 3 Acexamic acid (AK) 2.27 2.49 2.75 Aminoacetic acid 0.89 0.10 0.11 Sorbitol 1.96 2.17 2.40 Sodium Cyclamate 0.17 0.19 0.22 Fragrance food Orange 9374240 0.019 0,021 0,022

The proposed ratio of active substance and target additives was found experimentally and is optimal, ensuring AK powder meets all the requirements of the State Pharmacopoeia XI ed. and shelf life of at least 2 years (table 2).

table 2 №№ Name of quality indicator Quality Standards Actual figures Example 1 Example 2 Example 3 one. Description The contents of the package are white or white with a yellowish or creamy tint powder with the smell of an orange. powder with a smell of orange, white with a yellowish shade. orange powder orange powder with a creamy tint 2. Dissolution time The dissolution time of the contents of the package in 50 ml of warm water is not more than 2 minutes 1.8 1,5 1.9 3. The average weight of the contents of the package 4.85 to 5.15 g 4.98 5.01 5.09

four. pH 2.5 to 3.5 2,8 3.0 3.2 5. Mass loss on drying No more than 0.5% 0.16 0.23 0.21 6. Foreign matter Any impurity no more than 0.5%, total no more than 1.0% 0.3 / 0.81 0.21 / 0.60 0.29 / 0.59 7 Shelf life 2 years 2 years 2 years 2 years

Pharmacological tests of this dosage form were conducted (hereinafter this powder - the contents of the package for brevity will be referred to simply as AK).

MATERIALS AND METHODS

The experiment was performed on 45 white rats (males, weight about 150 g). Animals were divided into 3 series:

1 series - control, animals with experimental fracture of the tibia;

2 series - animals with experimental fracture of the tibia, which received 1.0 ml of placebo twice a day;

Series 3 - animals with an experimental fracture of the tibia, which were given orally twice a day with AK in a solution of 1 ml - 15 animals;

The therapeutic dose for rats was 0.25 g / kg.

Animals were kept in a vivarium of 5-6 rats in one cage. Bone tissue regeneration in rats was studied after the application of a defect in the diaphysis region of the right tibia. The operation was carried out under ether anesthesia under sterile conditions. A linear skin incision was made on the medial surface of the right lower leg. Muscles were separated from the tibia in a blunt manner. With special forceps, the exposed bone broke in the middle third. At the end of the operation, tightening sutures were applied to the skin. Under these conditions, relatively uniform fractures with a slight displacement of fragments were obtained, because the fibula played the role of a natural splint.

The collection of material for histological studies and radiography was carried out after 20, 30 and 40 days. Our judgments about the speed and nature of the reparative process were based on a comparison of macro-, microscopic, and x-ray studies of the fracture area. During osteogenesis, animals were removed from the experiment by an overdose of ether anesthesia. The area of injury was carefully monitored. After examining the damaged limb, its radiography was performed.

After radiography, the fracture area was exposed, checked for the mobility of the fragments, the density and size of the formed corns. For histological examination, a section of the tibia containing corn was excised. The material was fixed in 10% neutral formalin buffered according to Lily, Carnoy fluid, 96 °, 80 ° and 70 ° alcohols. Decalcification was carried out in 10% formic acid, a mixture of 40% formic acid with sodium nitrate (1: 1). After decalcification, bone sections were carried out using alcohols and contained in paraffin. Longitudinal histological sections were made, the preparations were stained with hematoxylin and eosin according to the method of Van Gieson and Mallory.

RESEARCH RESULTS.

X-RAY AND MICROSCOPIC CHARACTERISTICS OF THE PROCESS OF CONSOLIDATION OF Fractures IN THE CONDITIONS OF INTRODUCTION AK.

The results obtained during x-ray and microscopic studies of post-traumatic regeneration of bone tissue are the foundation for constructing our conclusions about the speed and completeness of the consolidation process.

In this regard, the study of the features of reparative osteogenesis that develops when AK is administered to experimental animals, we considered it necessary to begin with an analysis of the x-ray picture of corn formation.

Fracture healing was observed in dynamics over 40 days.

In control animals, 20 days after the injury, the development of a periosteal reaction is revealed, expressed more actively from the side of the distal fragment. The shadow from the forming callus is well expressed, has a significant extension, a clear, even contour. The interfragment space is presented in the form of a gear gap.

Partial fusion of fragments in the control can be noted after 30 days. A complete formation of the periosteal clutch occurs, which covers the ends of the fragments and enters the medial surface of the proximal fragment and the lateral surface of the distal fragment. A gradual reconstruction of the periosteal callus begins and after 40 days the ends of the fragments are fused with primary bone callus, which has the appearance of a homogeneous structure. The medullary canals at the ends of the fragments are still closed and do not communicate with each other.

In animals that received a placebo daily, the x-ray picture at all periods did not differ much from the images of rats in the control series.

In the third experimental series in animals treated with AK, there is a significant increase in the intensity of reparative osteogenesis.

After 20 days, the bone fragments are fused together with a well-developed periosteal callus, which in the form of a dense homogeneous shadow covers the regenerate area and extends far to the ends of the bone fragments. In most animals, the formation of primary bone marrow occurs, clutch-like fastening both bone fragments. The corn has a clear and smooth contours, uniform in structure. The cortical layer and the medullary canal are visible. On the 30th day of the experiment, all signs of the formation of cancellous bone tissue are found, and the fracture line is no longer traced.

After 40 days, the area of the former defect can only be determined by the wider growth of bone tissue from the side of the periosteum and a certain narrowing of the medullary canal in the fracture zone.

Thus, the consolidation of the proximal and distal fragments of the bone with complete restoration of the bone marrow canal is revealed radiologically.

During histological study of bone regenerate tissue preparations, 20 days after the fracture, distinct differences were observed in the animals of the compared experimental series (Table 3).

Table number 3 INFLUENCE OF AK ON THE DEVELOPMENT OF MOBILE BONE TISSUE DURING BONE REENERATION Series Res. whether Width of bone-beam structures (μm) 20 days 30 days 1 series (control) M ± m 36.3 57.1 1,08 2.31 2 series (fracture + placebo) M ± m 37.8 58.2 2.21 2.65 3 series (fracture + AK) M ± m 49.2 68.9 1.97 3.01

In the control at this stage, well-defined peri-and endo-osteal corns are noted. The newly formed osteoid tissue occupies the entire regenerate area along the fragments and reaches the fracture line. Trabeculae form loops surrounded at the periphery by layers of osteoblasts. The resulting beams are disoriented and have a significant thickness (table. No. 3). The inter-fragment space is performed by cartilage and chondroid tissue, between the fields of which there are strands of connective tissue. Intermediately are chondroid and areas of granulation tissue. Endo-osteal reaction manifests itself in the formation of young bone beams filling the lumen of the medullary canal.

Table number 4 NUMBER OF OSTEOCYTES PER UNIT OF BEAM AREA AT DIFFERENT STAGES OF OSTEOGENESIS (IN 100 FIELDS OF VISION) Series The timing 20 days 30 days 40 days 1 series (control) 184 ± 3 158 ± 2 131 ± 2 2 series (fracture + placebo) 179 ± 2 154 ± 3 127 ± 3 3 series (fracture + AK) 161 ± 2 124 ± 3 83 ± 2

In animals receiving a placebo, the histological picture of the fracture site does not differ significantly compared to rats in the control group. Osteoid tissue also performs the entire area of damage. The number of osteocytes per unit area is 179 ± 2 (table. No. 4). The regenerate is still heterogeneous, chondroid tissues border on connective and granulation. The thickness of the bone beams does not significantly differ from the control indicators (table. No. 3). The medullary canal is filled with newly formed bone beams.

Histological examination of tissues in animals treated with AK reveals a simultaneous active reaction in all areas of the regenerate. 20 days after the injury, rapid development of young spongy tissue is observed. The formed peri-and endo-ostal beams are oriented along the bone length (table. No. 3). Among the trabeculae, there are individual large, intensely stained, vacuolated cartilage cells with a pycnotized nucleus and cytoplasm. Proliferating osteoblasts in layers are located on the edges of the bone beams. Cartilage tissue is reformed and virtually the entire area of damage is filled with spongy bone tissue. Young bone beams in the central part of the corns and periosteal trabeculae acquire signs of lamellar structure, their further thickening occurs (49.2 μm versus 36.3 in the control) and a decrease in the number of cells contained in them (161 ± 2). In the central part of the corn, a portion of cartilage tissue penetrated by connective tissue fibers is still preserved. Endo-ostally, it is also possible to mark the beginning of the transformation of coarse-fibrous tissue into lamellar. The osteoblastic reaction continues to remain quite active (table. No. 4).

The following study of microscopic changes in the area of the bone defect was carried out 30 days after the fracture.

In control rats, the area of the bone defect is represented mainly by coarse fibrous bone tissue, which is formed both from the side of the periosteum and endoost, as well as intermediately. Bone cells are clearly identified, the color of the beam structures is uneven, the inter-beam spaces are filled with osteoblasts and connective tissue cell elements. A strip of cartilage and connective tissue passes through the central part of the whole corn. The lumen of the medullary canal near the lesion area is filled with pericladins, between which is located the yellow bone marrow.

In animals treated with placebo, no significant differences were found during the reparative process in the fracture zone (table. No. 3 and table. No. 4). As in the control, the defect area is filled with newly formed bone tissue with a distinct strip of cartilage and connective tissue in the center.

In a series of animals treated with AK, in contrast to the control, fragments are fastened with more massive bone tissue, the newly formed structures are more mature, which is confirmed by a decrease in the number of osteocytes in them (124 ± 3 versus 158 ± 2 in the control). Formed beams acquire signs of plate structure, mainly they are oriented along the bone. The bone beams that form the periosteal callus have a coarse-grained pattern. Their width is 68.9 ± 3.01 μm. In parallel with the restructuring into the lamellar bone, an intensive process of resorption of bone beams and the formation of cavities of the yellow and myeloid bone marrow occur. Endoostal bone callus in animals of this series is formed simultaneously with periosteal. The newly formed beams also acquire signs of compaction, and some of them are absorbed. The central zone of the defect under conditions of AK exposure undergoes active resorption and the formation of bone marrow hematopoiesis in the formed cavities. The increased vascularization of this regenerate zone is noteworthy.

After 40 days from the beginning of the reparative process, we revealed the following histological changes.

In control rats, the bone bonding corn somewhat decreases in size compared with the previous study period. Fragments are welded together by ripe spongy tissue, an active transformation of coarse-fibrous tissue into lamellar is underway. The trabeculae of the periosteal callus are oriented along the fragments, thicken, have a large-loop nature, compared with the previous period, a decrease in the number of osteocytes immured in them is noted (131 ± 2). Osteoblasts are arranged in rows along the beams. The total number of bone beams is reduced, inter-beam spaces are filled with yellow and myeloid bone marrow. Gradually, a new cortical layer begins to form from the intact bone.

When the organism of experimental animals AK was saturated, after 40 days from the beginning of the reparative process, we noted the formation of a more perfect callus, compared with the control. The thickening of the diaphysis at the fracture site is almost completely smoothed out. The periosteum is restored throughout. The cortical layer of the intact areas of the bone gradually passes into the young layer. Partially near the intact bone, bone tissue is compacted. In the periosteal zone, massive beams merge and form a compact substance with extensive inter-beam spaces and Haversian channels of primary osteons. In the bone marrow canal, thinning of the beams occurs, and in the inter-beam spaces the cell and yellow marrow are formed. The number of osteocytes naturally decreases and is 83 ± 2 against 131 ± 2 in the control. Simultaneously with osteoblasts, multinuclear large osteoclasts appear, which determine the sites of bone formation and destruction in different zones of the regenerate. Endosteal callus is reduced.

CONCLUSION

The results of a comprehensive phased study of post-traumatic regeneration of bone tissue, carried out using x-ray and histological techniques, allowed us to establish that:

1. With the oral administration of AK to experimental animals, the individual stages of the process of osteogenesis are significantly accelerated while maintaining the sequence of their passage. Under these conditions, the qualitative and quantitative characteristics of the emerging regenerate structures change significantly.

2. High proliferative activity of mesenchymal cells is noted, their accelerated differentiation into fibroblasts and osteoblasts producing a significant amount of fibrous connective and osteoid tissues in the composition of periosteal and provisional corns.

3. Under the conditions of AK administration during reparative osteogenesis, equally intensive formation of periosteal, intermedial, and endoostal corns occurs at different stages of the bone formation process.

4. Accelerated formation of the cancellous bone, consisting of wide bony beams with a large-loop pattern, is combined with the active transformation of coarse fiber into lamellar tissue, which is associated with the rapid restoration and intensive germination of regenerated tissues by blood capillaries, the appearance of primary crystallization nuclei and subsequent salt deposition, which is significantly shortens the final healing time.

In addition, the technology for the preparation of the proposed dosage form is simple and also convenient for use by patients.

Claims (2)

1. A solid dosage pharmaceutical composition containing acexamic acid, with the ability to reduce exudation, accelerate the cleansing of the wound from necrotic masses, and accelerate tissue epithelization and regeneration, characterized in that it contains acexamic acid, aminoacetic acid, sorbitol, sodium cyclamate, food flavoring Orange 9374240 in the following ratio of components, wt.%:
Acexamic acid (AK) 45.0-55.0 Aminoacetic acid 1.80-2.20 Sorbitol 39.24-47.96 Sodium Cyclamate 3.60-4.40 Fragrance food Orange 9374240 0.36-0.44 2. The pharmaceutical composition according to claim 1, characterized in that it is made in the form of a sachet.