BR102014029309B1 - INHALATION CHAMBER FOR IN VIVO BIOLOGICAL ASSAYS - Google Patents

Abstract

INHALATION CHAMBER FOR IN VIVO BIOLOGICAL TESTS, particularly a chamber intended to enable in vivo pharmacological and toxicological studies of vaporized and/or nebulized substances. For this, pressure control is not required, on the contrary, the chamber object of the invention has holes for the exit of the air inside it, allowing the renewal of this air without changing the pressure inside it. More specifically, this chamber aims to evaluate the therapeutic activity and toxicity of formulations for administration via the pulmonary route, in an in vivo model for rodents. In addition, it can also be used to assess the harm caused to the body by vapors, gases and fumes. The invention contemplates an inhalation chamber (1) with at least five "female" (2) recipients of the glass tubes (3) for packaging the animals, said glass tubes (3) each receiving a lid (4), and at the end of the inhalation chamber itself (1) is coupled the head? (5), while this inhalation chamber itself (1) has an air release area (6) close to its bottom; the referred ?head? (5) is coupled to a silicone hose (7) and this to a cylindrical glass joint (8) which is coupled at the opposite end to the hose (...).

Description

BRIEF SUMMARY OF THE INVENTION

[0001] This application for a Patent of Invention deals with an INHALATION CHAMBER FOR IN VIVO BIOLOGICAL TESTS, particularly a chamber that aims to enable in vivo pharmacological and toxicological studies of vaporized and/or nebulized substances. For this, pressure control is not required, on the contrary, the chamber object of the invention has holes for the exit of the air inside it, allowing the renewal of this air without changing the pressure inside it. More specifically, this chamber aims to evaluate the therapeutic activity and toxicity of formulations for administration via the pulmonary route, in an in vivo model for rodents. In addition, it can also be used to assess the harm caused to the body by vapors, gases and fumes.

FIELD OF THE INVENTION

[0002] The invention has particular application in the segment of in vivo biological assays, in the pharmaceutical, environment and smoking segments.

BACKGROUND OF THE INVENTION

[0003] Document BRMU8202611-4 is known, referring to a HYPERBARIC CHAMBER FOR USE IN EXPERIMENTS ON ANIMALS, deposited on 12/30/2002, which performs experiments under pressure control, the same occurring with document BRMU8300606-0 , of 05/20/2003, entitled HYPERBARIC CHAMBER. Therefore, they are technically outside the scope of this Invention Patent application.

[0004] Document US 4,305,347, published on 05/20/2003, refers to a TOXICOLOGICAL INHALATION CHAMBER, which comprises an inhalation chamber that works with a hermetic environment isolated from the external environment, which is not the purpose of this present invention patent application. In this previous document, the aforementioned hermetic environment does not mimic what happens when a medication is administered by inhalation via vaporization or nebulization, since, in this case, there is loss of part of the administered material and also constant renewal of it. In the construction shown in this previous document, four floors are contemplated, each one containing perforated cages in which the animals are, which can generate a differentiated distribution of the indoor air depending on its density and diffusion capacity, since the air will circulate through the lateral area .

[0005] An Inhalation Apparatus for testing on animals, associated with the concepts of the document in the previous paragraph, comprises the possibility of different doses of air being made available to animals because they are at different levels, as described in the chamber of the previous document, in addition to this apparatus presenting a bidirectional air flow inside the cylinders in which the animals are. Such an aspect can make it difficult to renew the air inside these cylinders, changing the dose of what is being evaluated and forcing the installation of the very odors that remain in circulation.

[0006] Document US 5,626,130, published on 06/05/1997 refers to a NASAL BREATHING DEVICE AND SYSTEM FOR LABORATORY ANIMALS, which, in summary, has the same purpose as the chamber of the present invention. However, the chamber of this previous document is not completely dismountable, which makes its handling, washing and use of cylinders with different diameters for different species of animals difficult. In this anterior chamber, the air is not homogenized inside the material to be tested, which makes dosage errors possible. The cylinders of this previous document are not of variable diameters, therefore, they are not able to accommodate the entire body of the animal, not guaranteeing frontal alignment with the air flow. Finally, this chamber in the previous document does not have a place to collect the air inside it for chemical evaluation.

[0007] Document WO 2010/002331, published on 01/07/2010, refers to an APPARATUS AND METHOD FOR EXPOSING LABORATORY ANIMALS TO A TEST SUBSTANCE, describes the homogenization of air with the substance to be tested inside the inhalation chamber. It is a fact that the homogenization inside the chamber can favor a heterogeneous distribution of the material to be tested for each of the animals under study, generating an analysis error. On the other hand, the anterior apparatus does not have an outlet at the end of the tubes where the animals are located, preventing the air from having a unidirectional flow, causing the inhalation of odors and gases from the animal itself, in addition to making it difficult to renew the air. to be inhaled and, consequently, of the material to be tested.

[0008] Documents CN 201684042 and CN 203029422 are also known, respectively dated 12/29/2010 and 07/03/2013, in which, as a common feature, it can be highlighted the fact that the animals are not always kept in the same line, which causes a difference in height and, as a result, a difference in the concentration and amount of inhaled air.

ANALYZING THE INVENTION IN RELATION TO THE STATE OF THE TECHNIQUE

[0009] This application for a Patent of Invention is notoriously different from the documents identified as prior art. In relation to documents BRMU8202611-4 and BRMU8300606-0, the invention is conceptually totally different, considering that it is not a hyperbaric chamber.

[0010] In relation to document US 4,305,347, the invention stands out due to the fact that the air released at the bottom of the chamber generates swirling and uniform diffusion through the cylinders arranged at the same height. Additionally, in the chamber of the present invention, the animals are placed in such a way that their snouts are aligned with the air entrance hole coming from inside the chamber, mimicking the tools used during inhalation. Another advantage of the chamber of the present invention consists of its size, obtained through the unprecedented construction, through which it is possible to facilitate transport, handling and washing, also presenting the possibility of working with a representative number of animals. Another important feature resides in the air outlet located in the final portion of the cylinder where the animals are located, which provides a constant renewal of the air inside these cylinders, in addition to enabling the elimination of odors from the animals being tested.

[0011] With regard to document US 5,626,130, the chamber object of the present invention stands out for being completely dismountable, which facilitates its handling, washing and use of cylinders with different diameters for different species of animals. On the other hand, in the chamber of the present invention, the air is homogenized inside the material to be tested, which minimizes dosage errors and mimics what happens in the clinic. As a result, the air internalized at the bottom of the chamber undergoes turbulence and diffuses homogeneously and unidirectionally to the cylinders containing the animals, that is, the air that reaches the cylinder will be inhaled and its excess along with residual vapors from the animals will be eliminated through the rear hole of the chamber. Furthermore, in the chamber of the present invention, there is no need to adapt new segments to accommodate the animal during the test, given that the cylinders of the chamber for which the invention is required can have variable diameters, therefore these cylinders are able to accommodate the entire body of the animal, keeping said animal always frontally aligned with the air flow.

[0012] In relation to document US 2009/0211534, the chamber of the present invention is not limited only to the use with aerosol, nebulization and vaporization systems of drugs and active substances can be adapted, in addition to toxicological tests for harmful fumes and vapors .

[0013] Dealing specifically with document WO 2010/002331, the chamber of this invention allows the material to be tested to be added through prior homogenization, since the main purpose is to mimic what happens in practice when we inhale something from nebulization, vaporization or combustion. From the moment that the already homogenized material is added to the chamber, the animals will inhale homogeneous ambient air at all points. In the chamber of the invention, the presence of extractor materials in said chamber can favor the adsorption of volatile substances essential for the therapeutic activity of the material to be tested, therefore the opening at the end of the tube where the animals are allows the elimination of volatile or suspended residues in the indoor air.

[0014] In relation to the Chinese documents CN 203029422 and CN 201684042, the chamber of the present invention stands out for keeping the animals always in the same line, that is, there is no difference in height, therefore, there will be no difference in concentration or quantity of air inhaled by influence of the density or renewal of that air.

[0015] Therefore, in relation to all prior art documents listed here, the chamber of the present invention is innovative in that it has a hole in the back of the cylinders in which the animals are located. As this is the only air outlet, the flow is forced through the animals due to physical pressure differences between the inside of the chamber and the outside environment. In this way, the air is breathed by the animals, being constantly renewed and still functioning as a carrier of volatile or suspended products from the animals.

ADVANTAGES OF THE INVENTION

[0016] Some of the improvements of the chamber developed in relation to the devices found in the literature are: - Decrease of the internal volume. - Need to use only one nebulizer device or any other steam source. - Air nebulized at the bottom of the chamber, central part, generating turbulence and uniform distribution to the tubes where the animals are. - Tubes where animals are removable, fitted by ground thread, facilitating washing and reducing the risk of breaking the glass apparatus. - Narrowing of the animals' tubes, preventing them from changing direction inside. - Lids of the tubes where the animals are kept, fitted with a ground joint and secured with an elastic band, containing an air outlet hole at the back, causing the nebulized air to pass through the animals to leave the chamber. - The unidirectional air outlet prevents the presence of contaminants inside the inhalation chamber. - Use only of glass parts that can be washed and sterilized so that there are no residues and contaminants that interfere with the tests, while the small silicone hose and the septum of the vial are replaced, with the same purpose, at each new analysis .

DESCRIPTION OF THE DRAWINGS

[0017] The invention will be explained below in a preferred form of embodiment, and, for a better understanding, references will be made to the attached drawings, in which they are represented: FIG. 1: Shows, in perspective, a representation of the central body of the inhalation chamber with its structures. FIG. 2: Shows, in perspective, a representation of the animals' packaging tube during the tests. FIG. 3: Shows, in perspective, a representation of the coupling between the tube for storing the animals and the lid that prevents the aforementioned animals from escaping. FIG. 4: Shows, in perspective, a representation of the lids for the tubes used to keep the animals. FIG. 5: Shows, in perspective, a representation of the 'head', the central piece for capturing and distributing the air inside the chamber, with its structures. FIG. 6A: Shows, in perspective, a representation of the coupling between the central body of the camera and the ‘head’. FIG. 6B: Shows, in perspective, the air release area, near the bottom of the chamber to generate turbulence. FIG. 7: Shows, in perspective, a representation of the coupling between the glass junction, the silicone hose and the ‘head’ responsible for admitting air into the chamber. FIG. 8A: Shows, in perspective, a representation of the glass joint. FIG. 8B: Shows, in perspective, the coupling of the glass joint to the silicone hose. FIG. 8C: Shows, in perspective, the connection to the silicone hose and the nebulizer cup. FIG. 9: Shows, in perspective, a representation of the air flow, by means of indicative arrows, into the inhalation chamber and its structures, with the chamber completely assembled and coupled to a nebulization device. FIG. 10A: Shows, in perspective, a representation of the inhalation chamber in activity with mice. FIG. 10B: Shows, in perspective, a representation of the glass tube with a lid with the mouse inside. FIG. 11: Shows, in side view, the central body of the inhalation chamber. FIG. 12: Shows, in side view, the ‘head’ responsible for admitting air into the inhalation chamber. FIG. 13: Shows, in side view, the glass junction. FIG. 14: Shows, in side view, the glass tube for storing the animals. FIG. 15: Shows, in side view, the lid of the animals' tube. FIG. 16: Shows, in side view, a representation of all the parts that make up the inhalation chamber of the invention, fitted to each other. DESCRIPTION OF THE INVENTION

[0018] THE INHALATION CHAMBER FOR IN VIVO BIOLOGICAL TESTS, object of this patent application, comprises a chamber composed of an inhalation chamber itself (1) containing a series of structures, said inhalation chamber itself (1) configuring at least five 'female' ground joints (2) for receiving the glass tubes (3) for the animals' packaging, said glass tubes (3) each receiving a lid (4) that prevents the aforementioned animals from escaping, and, at the end of the inhalation chamber itself (1), the 'head' (5) is attached, which is a central piece for capturing and distributing air inside the chamber itself (1), with its structures, while this inhalation chamber itself (1) has an air release area (6) close to its bottom, to generate turbulence; said 'head' (5) is coupled to a silicone hose (7) and this to a cylindrical glass joint (8) which is coupled, at the opposite end to the silicone hose (7), to the nebulization cup (9 ) and this to the nebulization device (10).

[0019] The inhalation chamber itself (1) comprises rounded walls (11) with a reduced central internal space (12) in order to reduce the dissipation of the admitted air, which results in a greater use of the material tested due to the rapid saturation of the chamber itself (1). The central body (13) of the inhalation chamber was made with an Erlenmeyer flask, in which, in addition to its 'female' ground joint (14) in the upper portion, at least five lateral outlets with 'female' ground joints (2) were added to adaptation of the glass tubes (3), each containing two lateral hooks (15) for attaching elastics (E) intended to prevent the lids (4) of the tubes (3) for packaging the animals from loosening. In addition, the central body (13) has an outlet (16) for collecting and analyzing the air inside it, said outlet equipped with a thread (16B) to receive a threaded plastic cap (17) with a perforable septum (18) made in rubber, silicone, teflon, or other flexible material to avoid loss of ambient air during in vivo assays (Figure 1).

[0020] More specifically, according to FIG. 1, the inhalation chamber itself (1) comprises, in addition to the aforementioned components, an upper outlet edge (19) which succeeds the upper 'female' ground joint (14), from which the smooth glass wall of the "neck" extends. ” (20) of the inhalation chamber (1), forming the smooth convex glass body (21), while the 'female' ground joints (2) each have a lateral outlet (2B), 'female' ground joint ' side (2C) and side exit edge (2D).

[0021] In each of the at least five lateral 'female' ground joints (2) are coupled, as already mentioned, the glass tubes (3) in which the animals used in the tests are placed (Figure 2). These tubes can vary in diameter according to the size of the animals used or the tests carried out, always preventing the animals from changing direction and, consequently, preventing them from receiving air directly into their muzzle. Its structure is cylindrical, with a funnel (3B) at the coupling end to the central body (13) of the inhalation chamber itself (1), in order to allow the snouts of the tested animals to fit, resulting in direct administration of the air admitted into this chamber. region of the animal's body. At that end there is, in the glass tube (3), a 'male' ground joint (3C) to fit in the central body (13), preventing the loss of air into the environment. At its opposite end, this glass tube (3) has another 'male' ground joint (3D), this one having the same diameter as the tube itself, necessary for coupling the cover (4). This wide outlet in the opposite part (distal part of the chamber) of the tube is important to enable the animal's conditioning during the test preparation. In each of the glass tubes (3) the lids (4) are placed right after the animals are packed (Figure 3). This structure prevents the animals from escaping the chamber during the inhalation test.

[0022] Each glass tube (3), in addition to the above components, contains an air inlet (22) from the central body (13) of the inhalation chamber itself (1), which is positioned next to the 'male' ground joint (3C) for coupling to the lateral outlet of the central body (13), followed by a funnel (23) to fit the animal's muzzle, and this one of a smooth glass body (24) of the animal's packaging cylinder, being finished with a 'male' ground joint (3D) for coupling to the cover (4), configuring an air outlet (25) at the bottom.

[0023] The cover (4) of the tube (3) for packaging the animals consists of a 'female' ground joint (26) at one end and an air outlet (27) to the outside of the inhalation chamber (1) at another said end, which can be sealed by means of a plastic cap (17) threaded with septum (18) (Figure 4).

[0024] This air outlet (27) is opened during the tests, causing the air admitted into the chamber to be forced out through it, since it is the only outlet for the inhalation chamber itself (1). This fact ensures that all air admitted is distributed from the central body (13) of the inhalation chamber itself (1) to the tubes (3) containing the animals and from there to the outside of the chamber, carrying, in a constant flow, body odors, excrements and gases expelled by the animals themselves. In addition to these structures, all lids (4) have two side hooks (15B) for elastic support, which will also be fixed by means of hooks (15) present in the 'female' ground joints of the central body (13) of the inhalation chamber itself. dictates (1), preventing the lid from loosening if the animal forces it. In addition, the lid (4) reveals an upper thread (28) for the plastic lid (17), and, immediately following the side hooks (15B), a smooth glass wall curve (29) extends, as well as as mentioned, a 'female' ground joint (26) for coupling to the tube (3) for packaging the animals, ending in a smooth glass edge (30) of the lid.

[0025] The upper 'female' ground joint of the central body (13) of the inhalation chamber itself (1) receives the coupling of the 'head' (5) of the inhalation chamber (1), this part responsible for receiving and distributing of air inside the inhalation chamber (1) (Figure 5). This part, or 'head' (5) is composed of an upper entrance (31) presenting rings (32) with diameters in progression of diameter for coupling of silicone hose (7). After the entrance, the smooth glass tube for the passage of the admitted air (33), forms a curve (34), followed by the 'male' ground joint (35), with adapter (36) of the ground joint, from where the tube is moves to the outlet (37) for coupling the 'head' (5) to the central body (13) of the actual inhalation chamber (1), avoiding the loss of admitted material. The central tube, through which the air passes, extends from the 'head' (5) to the bottom of the central body (13) of the inhalation chamber (1), releasing the air near the base of the inhalation chamber (1) and , consequently, generates a turmoil that optimizes the saturation and uniform distribution of the air for the animals (Figure 6).

[0026] The end of the silicone hose (7) opposite the one described is coupled to a cylindrical glass junction (8) equipped with a plurality of rings of different diameters (38), followed by this junction (8) of a tapering tube with a smooth glass wall (39), ending in a smooth glass joint (40) for coupling to the formulation container, where an inlet (41) of air from the nebulization cup (9) is configured, while the outlet (42) at the opposite end is air to the silicone hose (7) (Figures 7 and 8).

DESCRIPTION OF A PREFERRED CONSTRUCTION OF THE INVENTION

[0027] In the example shown in figure 9, it is observed that, from the compressor device (AC), the air is injected into the nebulization cup (9) containing the formulation, which, in turn, is nebulized and sent to the junction of glass (8) (Figure 8A). From this glass junction (8) the nebulized material passes through the silicone hose (7) (Figure 8B) and reaches the 'head' (5), the part responsible for admitting the nebulized air inside the inhalation chamber itself. (1) (Figure 5).

[0028] From then on, through the central tube of this piece, the nebulized air is introduced to the inhalation chamber (1) impacting with its bottom and generating a vortex (Figure 9), directing itself to the glass tubes (3 ) of animal conditioning (Figure 2). In this place, the animals inhale the material to be tested (formulations, vapors or fumes), while the residues (expired air, odors of the animal and its excretions) are carried to the outside of the inhalation chamber (1). The constant supply of air in the central body (13) means that the air in this environment is always renewed, since the newly arrived air forces the exit of the air already present in the chamber through the holes in the posterior region of the covers (4) belonging to the tubes (3) of conditioning the animals (Figure 3), avoiding supersaturation and contamination of the environment.

[0029] Figure 10 represents the inhalation chamber (1) being used for in vivo inhalation tests with mice. It is worth noting the presence of elastics (E) attached to the lid of the tube (3) for storing the animals and to the 'female' ground joint to prevent the animals from escaping. In addition, the turbidity inside the inhalation chamber (1) generated by the turbidity of the nebulized air inside the central body (13) can also be noticed.

Claims (7)

1) INHALATION CHAMBER FOR IN VIVO BIOLOGICAL TESTS, comprising at least five 'female' ground joints (2) for receiving the glass tubes (3) used for packaging the animals, said glass tubes (3) each receiving a lid ( 4); said 'head' (5) is coupled to a silicone hose (7) and this to a cylindrical glass joint (8) which is coupled, at the opposite end to the silicone hose (7), to the nebulization cup (9 ) and this to the nebulization device (10), characterized by having rounded walls (11) with a reduced central internal space (12), containing a central body (13) made with an Erlenmeyer flask, in which, in addition to its ground joint ' female' (14) in the upper portion, at least five side outlets with ground 'female' joints (2) were added to adapt the glass tubes (3) with respective covers (4), each containing two side hooks (15) ; the central body (13) has an outlet (16) provided with a thread (16B) to receive a threaded plastic cap (17) with a perforable septum (18) made of rubber, silicone, teflon or other flexible material. 2) INHALATION CHAMBER FOR BIOLOGICAL IN VIVO TESTS, according to claim 1, characterized in that it comprises an upper outlet edge (19) which succeeds the upper 'female' ground joint (14), from which the smooth glass wall extends of the "neck" (20), forming the smooth convex glass body (21), while the 'female' ground joints (2) each have a lateral outlet (2B), lateral 'female' ground joint ( 2C) and side exit edge (2D). 3) INHALATION CHAMBER FOR IN VIVO BIOLOGICAL TESTS, according to claim 1, characterized in that in each of the at least five lateral 'female' ground joints (2) glass tubes (3) are coupled, which have a variable diameter , and its structure is cylindrical, with a funnel (3B) at the coupling end to the central body (13); in the glass tube (3) there is a 'male' ground joint (3C) for fitting in the central body (13); at its opposite end, this glass tube (3) features another 'male' ground joint (3D), this one the same diameter as the tube itself, for coupling the cover (4). 4) INHALATION CHAMBER FOR BIOLOGICAL IN VIVO TESTS, according to claim 1, characterized in that each glass tube (3) contains an air inlet (22) from the central body (13), which is positioned next to the ground joint ' male' (3C) for coupling to the lateral outlet of the central body (13), followed by a funnel (23) to fit the animal's muzzle, and this of a smooth glass body (24) of the animal's packaging cylinder, being finished with a 'male' ground joint (3D) for coupling to the cover (4), configuring an air outlet (25) at the bottom. 5) INHALATION CHAMBER FOR IN VIVO BIOLOGICAL TESTS, according to claim 1, characterized in that the cover (4) of the tube (3) for packaging the animals consists of a 'female' ground joint (26) at one end and an air outlet (27) to the outside at the other said end, which can be sealed by means of a plastic cap (17) threaded with a septum (18); in addition to these structures, all covers (4) have two side hooks (15B) for elastic support, which will also be fixed by means of hooks (15) present in the 'female' ground joints of the central body (13); the cover (4) configures an upper thread (28) for the plastic cover (17), and, immediately following the side hooks (15B), a smooth glass wall curve (29) extends, as well as the aforementioned a 'female' ground joint (26) for coupling to the tube (3) for packaging the animals, ending in a smooth glass edge (30) of the cover. 6) INHALATION CHAMBER FOR IN VIVO BIOLOGICAL TESTS, according to claim 1, characterized in that the upper 'female' ground joint of the central body (13) receives the coupling of the 'head' (5); this piece, or 'head' (5) is composed of an upper entrance (31) presenting rings (32) with diameters in progression of diameter for coupling of silicone hose (7), and, after the entrance, the smooth glass for the passage of air admitted (33), forms a curve (34), followed by the ground joint 'male' (35), with adapter (36) of the ground joint, from where the tube moves to the outlet (37) for coupling the 'head' (5) to the central body (13); the central tube, through which the air passes, extends from the 'head' (5) to the bottom of the central body (13). 7) INHALATION CHAMBER FOR BIOLOGICAL IN VIVO TESTS, according to claim 1, characterized in that the end of the silicone hose (7) is coupled to a cylindrical glass junction (8) equipped with a plurality of rings of different diameters (38) , then this junction (8) of a funneling tube with a smooth glass wall (39), ending in a smooth glass joint (40) for coupling to the formulation container, where an inlet (41) of air from the nebulizing cup (9), while the outlet (42) at the opposite end is for air to the silicone hose (7).

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