Container, device for releasing carbon monoxide, treatment system, set for treating living cells, method for treating mammals, and carbon monoxide

Abstract

The present invention relates to a container (10) configured for insertion into a human and / or animal body, the container (10) including a housing (12) and a chamber (14) defined within the housing (12). The chamber (14) is configured to receive a principal substance (42) configured to selectively emit carbon monoxide when activated by an activating substance (44). The chamber (14) is surrounded by a wall (16) of the housing (12). The wall (16) is configured to be permeable to carbon monoxide emitted by the principal substance (42) and to allow carbon monoxide emitted by the principal substance (42) to be released from the container (10) to the environment. The container (10) is configured such that the activating substance (44) is introduced into the chamber (14) from outside the housing (12) with the principal substance (42) already disposed within the chamber (14). The invention further relates to a device (40) for selectively releasing carbon monoxide, a treatment system (60; 80) and a set for treating biological cells (preferably living cells), a method for treating mammals, uses of carbon monoxide, and a method for treating biological cells.

Description

[Technical Field]

[0001] The present invention relates to a container, a device for releasing carbon monoxide, a treatment system, a set for treating living cells, a method for treating mammals, and carbon monoxide. [Background technology]

[0002] A wide variety of substances, e.g., gases and / or liquids, are used for therapeutic purposes, e.g., to treat one or more disorders. Each substance may be applied externally, e.g., to the skin of a subject, e.g., a human and / or animal. However, in some cases, it may be necessary and / or more effective to introduce the substance into the subject's body.

[0003] In some cases, it may be advantageous to house each substance in a housing, such as a capsule, to facilitate administration of each substance to a treatment site, e.g., within a subject's body, such that the substance is released at the treatment site, e.g., within the subject's body, particularly when the capsule or other housing reaches a specific location, e.g., a target site, within the subject's body. This can enhance the therapeutic effect of the substance and / or prevent or at least limit adverse effects that the substance may have on one or more areas of the subject's body.

[0004] For example, carbon monoxide (CO) is known to have beneficial therapeutic effects in humans and / or animals. For example, when a subject experiences one or more stressors, the body may naturally increase its production of carbon monoxide in response to the stressors to induce or enhance one or more countermeasures and / or defense mechanisms. Therefore, exogenous supplementation of carbon monoxide as a medical substance can be expected to have a relatively high therapeutic effect.

[0005] However, administering carbon monoxide to each subject poses several challenges. For example, carbon monoxide is generally considered a toxic gas. In particular, when a subject is exposed to certain levels of carbon monoxide, for example, by inhaling it, the carbon monoxide can mix and bind with hemoglobin in the subject's blood. This can result in a decrease in the amount of oxygen delivered to body tissues and vital organs, such as the brain and heart, potentially causing reversible and / or irreversible damage to the human and / or animal body, and even death.

[0006] Thus, there is a need for a safe and / or controlled method of applying carbon monoxide. Carbon monoxide release systems have been proposed in the prior art.

[0007] For example, European Patent Publication No. 3 242 552 B1 (incorporated herein by reference in its entirety) discloses a gas releasing device comprising two compartments. One of these compartments contains carbon monoxide releasing molecules (CORM), and the other compartment contains an activator that triggers the release of carbon monoxide from the CORM. When the substances from both compartments combine, a chemical reaction is initiated with the release of carbon monoxide. Both compartments are surrounded by a gas-permeable membrane that allows carbon monoxide to pass through. This membrane is impermeable to the CORM, the activator, and the reaction by-products.

[0008] Furthermore, International Publication No. 2021 / 074159 A1 (the entirety of which is incorporated herein by reference) describes a membrane-based gas-releasing device for oral administration. The device includes two compartments separated by an internal partition and surrounded by a gas-permeable membrane. One compartment is filled with carbon monoxide-releasing molecules, and the other compartment contains an activator that triggers the release of carbon monoxide from the carbon monoxide-releasing molecules. One compartment includes a hollow, tapered open end that tapers obliquely. When the device is squeezed to rupture the internal partition, this portion moves toward the internal partition, causing the carbon monoxide-releasing molecules and the activator to mix and release carbon monoxide through the gas-permeable membrane.

[0009] Furthermore, WO 2021 / 180908 A1 (incorporated herein by reference in its entirety) discloses a therapeutic system configured for the external treatment of wounds, inflammatory diseases of the skin, and inflammatory diseases of subcutaneous skin tissue, joints, and tendons, which includes a carbon monoxide-releasing molecule and a reaction chamber including a gas-permeable, liquid- and solid-impermeable membrane, in which the carbon monoxide-releasing molecule can be mixed with a carbon monoxide release-inducing compound.

[0010] However, there are several drawbacks associated with carbon monoxide emitting devices known in the prior art. For example, the configuration of carbon monoxide emitting devices known in the prior art, such as the two-compartment structure of the device disclosed in WO 2021 / 074159 A1, is relatively complex, resulting in a relatively large size and / or potential defects. Furthermore, due to the relatively complex configuration, each carbon monoxide emitting device known in the prior art is relatively expensive to manufacture, takes a relatively long time to manufacture, and / or has a relatively high risk of failure. Furthermore, the system described in WO 2021 / 180908 A1 is specifically designed to treat the external surface of a subject, particularly the subject's skin. For this purpose, the system described in WO 2021 / 180908 A1 includes a relatively complex mixture of materials and a gas-impermeable portion for directing the released carbon monoxide to the skin. Therefore, this system lacks flexibility for broader applications.

[0011] The application of carbon monoxide to various diseases and its signaling mechanisms are discussed, for example, in Gibbons et al., Carbon monoxide in gastrointestinal physiology and its potential in therapeutics, Aliment Pharmacol Ther. 2013 Oct; 38(7): 689-702. Summary of the Invention [Problem to be solved by the invention]

[0012] Thus, it is an object of the present invention to provide an improved means for applying carbon monoxide to a target. [Means for solving the problem]

[0013] A first aspect of the present disclosure relates to a container. The container may be configured to be inserted into a human and / or animal body, preferably via one or more natural orifices in the respective body. The container may include at least one housing and at least one chamber defined within the housing. The chamber may be configured to receive and store at least one principal substance configured to selectively emit carbon monoxide. The principal substance may be configured to emit carbon monoxide upon activation by at least one activating substance. The principal substance may be configured to emit carbon monoxide substantially immediately after the principal substance and the activating substance are mixed. Alternatively, the principal substance may be configured to emit carbon monoxide in a time-delayed manner, i.e., after a certain period of time has elapsed after the principal substance and the activating substance are mixed. If the principal substance emits carbon monoxide as a result of a reaction between the principal substance and the activating substance, the reaction may be heat-dependent. For example, the principal substance may begin emitting carbon monoxide when the mixture of the principal substance and the activating substance is heated to a certain threshold temperature, e.g., by body temperature within the human and / or animal body.

[0014] The chamber may be enclosed by at least one wall of the housing, at least a portion of which may be at least partially permeable to carbon monoxide evolved from the principal substance, such that the container is configured to at least partially release carbon monoxide evolved from the principal substance to the environment.

[0015] The "environment" may be a treatment site, such as, for example, within a human and / or animal body, or may be within a container for receiving biological cells, preferably living cells, such as immune cells, e.g., CAR T cells and / or stem cells, and / or gametes, e.g., sperm and / or oocytes, in which the cells are treated with carbon monoxide released from the container.

[0016] The container may be configured to allow an activator to be introduced into the chamber from outside the housing, with the activator already disposed in the chamber, and may be configured such that when the activator is introduced into the chamber, the activator and the principal substance react through a wall of the housing to release carbon monoxide.

[0017] That is, in contrast to carbon monoxide releasing devices known in the prior art, the activating substance does not have to be contained in a container. Instead, the activating substance may be introduced into the chamber from outside the housing, with the principal substance already disposed in the chamber, i.e., with the principal substance sealed in the chamber, preferably substantially fluid-tight and / or air-tight, from the environment. In other words, the principal substance may be stored in the chamber until the container is used. Before the container is administered, the activating substance can be introduced into the chamber from outside the housing to activate the principal substance. This allows for a simplified container structure.

[0018] In particular, the active substance can be introduced into the chamber from outside the housing while the principal substance is already disposed therein, eliminating the need for a separate chamber in the container to accommodate the active substance before mixing them. That is, the container may have a relatively simple configuration, for example, including only one chamber for accommodating the principal substance, with the active substance being introduced directly into the chamber containing the principal substance, allowing the principal and active substances to be mixed within the chamber. Furthermore, in contrast to prior art carbon monoxide release devices, such as those disclosed in EP 3 242 552 B1 and WO 2021 / 074159 A1, one or more means for separating the active substance from the principal substance are not required, and such means can be omitted. This may also allow for a more compact container configuration compared to prior art carbon monoxide release devices, potentially facilitating application (e.g., ingestion) of the container.

[0019] Furthermore, as described above, since the structure of the container is simplified, the manufacturing of the container is also simplified, and the manufacturing labor of the container, for example, the manufacturing cost and / or manufacturing time, may be reduced. Furthermore, the manufacturing process for manufacturing the container may generally be less prone to defects.

[0020] Additionally, configuring the container so that an activating substance can be introduced into the chamber from outside the housing with the principal substance already disposed within the chamber may provide a relatively simple and / or reliable activation process for activating the carbon monoxide-releasing principal substance. That is, a relatively safe and / or relatively reliable carbon monoxide-releasing mechanism may be provided that can prevent, or at least reduce the risk of, potentially adverse effects of the carbon monoxide-releasing principal and activating substances on a subject treated by the container. The carbon monoxide-releasing substance and / or its corresponding activating substance may be toxic to the subject, for example, if the subject were exposed to the carbon monoxide-releasing substance and / or its respective activating substance due to, for example, a structural defect in the container and / or a malfunction of the container.

[0021] Additionally, configuring the container to be insertable into a human and / or animal body can broaden the range of potential applications of the container, thereby increasing the flexibility of its use. For example, the container can be configured for in vivo use by rectally and / or orally introducing the container into a subject's body. For example, if the entire container is configured as a capsule, such a capsule can be configured for insertion into one or more body cavities of an animal and / or human, such as for oral and / or rectal administration to an animal and / or human, e.g., a child and / or adult, and / or for insertion, preferably manually, into the ear of an animal and / or human for the treatment of, but not limited to, otitis media and / or other inflammatory conditions for which targeted release of CO may be useful in treating, and / or for insertion into the vagina of an animal and / or human.

[0022] Preferably, such a container is configured to be ingested, e.g., swallowed, by a human and / or animal. The container can be configured for insertion into a human and / or animal body, allowing for various applications. For example, such a container can be placed within a container containing one or more biological cells, e.g., immune cells such as CAR T cells and / or stem cells, and / or gametes, e.g., sperm or oocytes, to provide ex vivo therapeutic treatments that can therapeutically treat and / or prevent degradation of the biological cells. Preferably, such a container is configured for insertion into a human and / or animal body via one or more corresponding natural orifices in the body.

[0023] In contrast, the system described in WO 2021 / 180908 A1 is specifically designed to treat an external surface, such as the skin, of a subject. To this end, the system described in WO 2021 / 180908 A1 includes a relatively complex mixture of different materials and a gas-impermeable portion to direct released carbon monoxide to the subject's skin. The use of such a composition and mixture of different materials requires a relatively complex and expensive manufacturing process. Furthermore, the use of such a composition and mixture of different materials can make it difficult, or even impossible, to miniaturize the device to a smaller size so that it can be inserted into a human and / or animal body, for example, orally and / or rectally.

[0024] Preferably, the entire wall of the housing, e.g., the entire perimeter of the housing / container, is permeable to carbon monoxide emitted by the principal substance, which allows carbon monoxide emitted by the principal substance to escape from a relatively large surface of the container, which may in turn improve the therapeutic efficacy of the container.

[0025] Preferably, when an activating substance is introduced into the chamber from outside the housing with the principal substance already disposed in the chamber, the container is configured such that after introduction of the activating substance via an introduction device such as a hollow needle is completed, e.g., after the introduction device such as a hollow needle is withdrawn from the housing, a substantially liquid-tight and airtight seal is provided at the introduction path portion of the housing for the activating substance. In other words, the introduction path portion of the housing for the activating substance may be configured to be resealed after introduction of the activating substance is completed, e.g., after the introduction device such as a hollow needle is withdrawn from the housing.

[0026] Such a container may include one or more designated sections configured to enable and / or facilitate the introduction of an active substance into the chamber and / or to provide a substantially fluid-tight and air-tight seal after introduction of the active substance is complete and an introduction device, e.g., a hollow needle, is withdrawn from the housing. For example, the housing may include one or more designated sections configured to introduce the active substance into the chamber, to be pierced by an introduction device, preferably a hollow needle, and to reseal after the introduction device is removed from the chamber. One or more designated sections may have an increased amount of material, e.g., an increased thickness of material, compared to adjacent sections of the housing, e.g., to facilitate piercing the housing through the one or more designated sections and / or to facilitate resealing of the one or more designated sections after the respective introduction device is removed from the chamber.

[0027] The container may have a substantially smooth and / or rounded outer shape, in other words, the container may have no sharp edges along its outer shape, preferably along its entire outer shape, which may reduce discomfort and ease the subject's introduction of the container into the subject's body, for example by ingestion, e.g., swallowing, and / or inserting the container into the rectum.

[0028] The housing may be constructed as a single, integral part, such that, for example, the principal and activating substances can be introduced into the completely enclosed chamber by piercing a portion of the wall of the housing, e.g., with a hollow needle, and then injecting the principal and activating substances into the chamber. Alternatively, the housing may include multiple components, e.g., a body and a lid, which are assembled together.

[0029] The entire container is preferably made of a single material, such as silicone. Being made of a single material does not mean that the entire container is integrally constructed as a single part. Being made of a single material means that only a single material, such as silicone, is used in the manufacture of the container. This allows for less labor in the manufacture of the container and / or further simplifies the structure of the container. This also allows for, for example, reducing the size of the container to make it easier to adapt to each application.

[0030] The activating material can be selected from a variety of suitable materials. The activating material may be liquid and / or gaseous, at least at the time of application of the activating material to activate the carbon monoxide-releasing principal material. For example, examples of the activating material include FeCl, Ce(SO), or H,O, with FeCl being preferred.

[0031] The principal substance may be configured to emit carbon monoxide upon activation, such as by reaction with an activating substance. The carbon monoxide-releasing principal substance may be selected from a variety of suitable materials. The principal substance may be liquid and / or gaseous and / or solid at the time the activating substance is applied to activate the carbon monoxide-releasing principal substance. The carbon monoxide-releasing principal substance is preferably a metal carbonyl compound, more preferably a molybdenum carbonyl compound, and most preferably a salt form of MO(CO)3(CNCH2CO2H)3, preferably Na3Mo(CO)3(CNCH2CO2)3.

[0032] Although this disclosure focuses on the release of carbon monoxide from the container, the container may be configured to release any gas, particularly any type of gas that may be used for therapeutic purposes. The principal and activating substances may be selected accordingly to provide the desired gas, and the wall material of the housing may be selected accordingly to be at least partially permeable to the respective gas.

[0033] As noted above, the container preferably includes only a single chamber, and can be configured such that the principal substance reacts with the activator when the activator is inserted into the single chamber from outside the housing by introducing the activator into the single chamber from outside the housing with the principal substance already disposed in the chamber.

[0034] The walls of the enclosure may be impermeable to one or more of solids, liquids, toxic substances, preferably at least heavy metal atom-containing substances, and by-products, preferably at least toxic by-products, resulting from the reaction of the principal substance with the activator substance.

[0035] Such containers are preferably configured to release carbon monoxide in vivo and / or ex vivo. Preferably, such containers are configured to be inserted into one or more body cavities of an animal and / or human (e.g., a child and / or an adult), e.g., orally and / or rectally, and / or into the ear and / or vagina of the animal and / or human, for example (but not limited to) the treatment of otitis media or other inflammatory conditions in which targeted release of CO may be therapeutically useful. Preferably, such containers are configured to be ingested, e.g., swallowed, by the human and / or animal.

[0036] Preferably, at least one section of the housing is configured to be penetrated by an introduction device configured to introduce an activating substance into the chamber for activation of the principal substance, and the at least one section of the housing is configured to reseal, preferably in a substantially liquid-tight and / or gas-tight manner, after the introduction device is withdrawn from the housing. The introduction device may comprise a hollow needle, for example a syringe with a hollow needle.

[0037] The housing preferably includes at least one body and at least one lid. The body and lid are preferably configured to be connectable to one another, preferably in a substantially liquid-tight and / or gas-tight manner. The body and lid can be connected to one another by a friction-type connection, a form-locking connection, and / or a material adhesive connection. The material adhesive connection can be achieved, for example, by at least one adhesive, such as at least one silicone adhesive.

[0038] Preferably, the lid and / or body include a protrusion configured to project into the chamber when the lid and body are interconnected.

[0039] Preferably, the protrusion is configured as a barrier that can be penetrated by an introduction device configured to introduce an activating substance into the chamber to activate the principal substance. Preferably, the protrusion is configured to reseal in a substantially liquid-tight and / or air-tight manner when the introduction device is withdrawn from the protrusion. By configuring the barrier of the container configured to introduce an activating substance into the chamber as a protrusion that protrudes into the chamber rather than outwardly from the chamber, it is possible to prevent the protrusion from causing discomfort to the subject and / or interfering with application or administration of the container when inserting the container into the body of a subject, for example into the rectum and / or mouth.

[0040] For example, when the container is used ex vivo, e.g., when biological cells, preferably live cells, are processed within the container, a protrusion extending outward from the chamber can be disadvantageous. For example, a protrusion extending outward from the chamber can increase the overall dimensions of the container, making it more difficult to place the container in a relatively compact space, e.g., when the container contains live cells to be processed by carbon monoxide emitted from the container. Furthermore, by configuring the septum of the container configured to introduce an active substance into the chamber as a protrusion extending into the chamber, it is possible to prevent, or at least reduce the risk of, damaging the septum when the active substance is inserted into the chamber of the container, resulting in unintended release of the carbon monoxide-releasing substance and / or the active substance, and / or allowing one or more substances (e.g., body fluids, etc.) to enter the container through a damaged portion of the septum.

[0041] The protrusion may have any shape, preferably having a substantially circular cross-sectional shape when viewed perpendicular to the longitudinal axis extending into the chamber when the lid and body are connected together.

[0042] Preferably, the protrusion has a rounded or chamfered edge at the intersection of one or more side walls and one or more end faces of the protrusion, e.g., along the longitudinal axis of the housing, where the protrusion terminates within the chamber. This may facilitate assembly of the container. This feature may particularly facilitate assembly of the lid and body, i.e., by preventing or at least reducing the risk that a relatively sharp edge of the protrusion on the lid may catch on a surface of the lid or body during assembly of the lid and body, making assembly more difficult or impossible, and / or by preventing or at least reducing the risk that a portion of the container may be damaged during assembly of the lid and body.

[0043] The protrusion preferably has a length along the interior of the chamber and / or along the longitudinal axis of the lid and / or body when the body and lid are connected to one another, the length of the protrusion being at least 1 mm, more preferably at least 1.2 mm, more preferably at least 1.4 mm, more preferably at least 1.6 mm, more preferably at least 1.8 mm, more preferably at least 2 mm, more preferably at least 2.2 mm, more preferably at least 2.4 mm, more preferably at least 2.6 mm, more preferably at least 2.8 mm, more preferably at least 3 mm, more preferably at least 3.2 mm, more preferably at least 3.4 mm, more preferably at least 3.6 mm, more preferably at least 3.8 mm, more preferably at least 4 mm, more preferably at least 4.2 mm, more preferably at least 4.4 mm, more preferably at least 4.6 mm, more preferably at least 4.8 mm, more preferably at least 5 mm, more preferably at least 5.2 mm, more preferably at least 5.4 mm, more preferably at least 5.6 mm, more preferably at least 5.8 mm, and most preferably at least 6 mm. The protrusion length values ​​specified above can provide a balance between ensuring sufficient length for the activator to be introduced into the chamber, allowing the activator to be introduced into the chamber through a protrusion configured as a septum so that the introduction point of the protrusion reseals after the activator has been introduced into the chamber, while limiting the amount of space the protrusion takes up within the chamber.

[0044] The main body and / or lid preferably have a groove, which is preferably formed in a circular or semi-circular shape around the longitudinal axis of the main body and / or lid. In this case, it is preferable that the main body and the lid are connected to each other by at least partially inserting a portion of the main body or the lid that does not have a groove into the groove. This makes it possible to more reliably and firmly connect the lid and the main body. Furthermore, the groove also functions as a guide when assembling the lid and the main body, making it easier to assemble the lid and the main body. Furthermore, by configuring the main body and the lid so that the portion of the main body or the lid that does not have a groove is at least partially inserted into the groove when connecting the main body and the lid, it is possible to at least partially seal the gap between the main body and the lid.

[0045] Preferably, the protrusion defines a groove at least partially around the periphery of the protrusion when the lid and body are connected together, and preferably such groove is formed at least partially coaxially around the periphery of the protrusion.

[0046] Preferably the body and / or lid is made of silicone, preferably biocompatible silicone, preferably medical grade silicone.

[0047] Preferably, the entire lid is formed as a single piece. Alternatively, or in addition, the entire body may be formed as a single piece. This may facilitate the manufacturing process of the lid and / or the body. Alternatively, or in addition, it may facilitate assembly of the lid and the body. The lid and / or the body may each be molded and / or cast from a single material (e.g., silicone, etc.).

[0048] Preferably, the lid and / or the body has a Shore A hardness of 20 Shore A to 80 Shore A, preferably 30 Shore A to 50 Shore A.

[0049] To enhance sealing and / or friction between the body and lid when they are coupled together, the body and / or lid preferably include at least one sealing and / or friction-enhancing element disposed along at least one coupling interface between the body and lid. This can provide or increase the sealing and / or frictional force acting between the body and lid when they are coupled together. Alternatively or additionally, the sealing and / or friction-enhancing element can increase the engagement and / or adhesive area between the body and lid. This can reduce leakage from the container through the body-lid interface and / or reduce the risk of leakage. It can also prevent or at least reduce the risk of undesired or at least partial separation of the lid and body.

[0050] Preferably, the sealing element and / or friction-enhancing element includes at least one projection extending from the interface between the lid and / or the body. Such interface may extend along the connecting interface between the lid and the body. Such projection may be configured to project toward and engage the non-projecting portion of the lid or the body when the body and lid are connected together. Such projection may increase the force acting between the lid and the body, thereby improving the sealing effect between the lid and the body and / or increasing the connecting force between the lid and the body, thereby preventing or reducing the risk of undesired or at least partial separation between the lid and the body. Such projection may provide or increase a crimp between the lid and the body.

[0051] Preferably, the lid or body includes a protrusion. The interface of the non-protrusion-containing lid and body extending along the mating interface between the lid and body may be substantially smooth and / or continuous and / or non-protrusion-containing and / or non-dimple-containing, thereby increasing the force acting between the lid and body. Alternatively, the non-protrusion-containing lid or body may include at least one recess (e.g., at least one recess configured to at least partially receive a protrusion) and / or at least one protrusion.

[0052] Preferably, the protrusions project at least partially around the circumference of the boundary surface of the lid and / or body, preferably along the entire circumference, preferably multiple times, preferably in a spiral and / or thread-like and / or serpentine and / or zigzag manner, which may provide a relatively large surface area and thereby enhance the sealing and / or frictional forces of the protrusions. The circumference may be either the inner circumference and / or the outer circumference.

[0053] The projections are preferably formed from the same material as the rest of the lid and / or the protrusions and / or the body, and the projections and protrusions are preferably integrally and / or monolithically formed with their respective projections in a single moulding step when manufacturing the lid and / or the body.

[0054] Preferably, the lid includes a protrusion. The distal end of the protrusion, which penetrates deepest into the body when the lid and the body are coupled together, may have one or more rounded and / or chamfered edges. Alternatively, or in addition, at least a portion of the protrusion may be tapered toward the distal end of the protrusion, which penetrates deepest into the body when the lid and the body are coupled together. This may facilitate the removal of the molded lid from a mold and / or reduce the risk of damage during removal, compared to a cylindrical protrusion, for example, when the lid is manufactured by a molding process, thereby facilitating the manufacture of the lid. In addition, leaving a space or a larger space at the edge of the distal end of the protrusion, compared to a protrusion structured with one or more sharp edges at the distal end of the protrusion, may prevent or at least reduce spillage of a substance, e.g., an active substance, after removal of an introduction device, such as a hollow needle, for introducing the substance into the container.

[0055] Preferably, the maximum dimension of the entire container, preferably the length extending along the longitudinal axis of the container, is less than 25 mm, preferably less than 24 mm, more preferably less than 23 mm, more preferably less than 22 mm, more preferably less than 21 mm, more preferably less than 20 mm, more preferably less than 19 mm, more preferably less than 18 mm, more preferably less than 17 mm, more preferably less than 16 mm, more preferably less than 15 mm, more preferably less than 14 mm, more preferably less than 13 mm, and more preferably less than 12 mm. Configuring the container to be relatively compact in accordance with the above-specified maximum dimensions allows for easier introduction of the container into the human and / or animal body, particularly via one or more natural orifices therein, such as the mouth and / or rectum. Furthermore, providing a relatively small container allows for at least easier and / or more flexible placement within a relatively compact space to accommodate biological cells, preferably live cells, to be treated with carbon monoxide emitted from the container.

[0056] Preferably, the maximum dimension of the entire container in a direction extending substantially transverse to the longitudinal axis of the body and / or the longitudinal axis of the lid is less than 12 mm, more preferably less than 11 mm, more preferably less than 10 mm, more preferably less than 9 mm, more preferably less than 8 mm, more preferably less than 7 mm, more preferably less than 6 mm, more preferably less than 5 mm, more preferably less than 4 mm. As mentioned above, configuring the container to have relatively compact dimensions in a direction extending substantially transverse to the longitudinal axis of the body and / or the longitudinal axis of the lid may facilitate introduction of the container into the human and / or animal body, in particular via one or more natural orifices therein, for example orally and / or rectally, and / or may at least enable it to be more easily and / or flexibly positioned within a relatively compact space to accommodate biological cells, preferably live cells, to be treated with carbon monoxide emitted from the container.

[0057] The body preferably has at least one body wall that at least partially defines a chamber. The body wall is preferably at least partially permeable to carbon monoxide released by the principal substance activated by the activating substance. The body wall has a thickness of 0.4 mm to 2.5 mm, preferably 0.5 mm to 2.4 mm, more preferably 0.5 mm to 2.3 mm, more preferably 0.5 mm to 2.2 mm, more preferably 0.5 mm to 2.1 mm, more preferably 0.5 mm to 2 mm, more preferably 0.5 mm to 1.9 mm, more preferably 0.5 mm to 1.8 mm, more preferably 0.5 mm to 1.7 mm, more preferably 0.5 mm to 1.6 mm, more preferably 0.5 mm to 1.5 mm, more preferably 0.5 mm to 1.4 mm, more preferably 0.5 mm to 1.3 mm, more preferably 0.5 mm to 1.2 mm, more preferably 0.5 mm to 1.1 mm, more preferably 0.5 mm to 1 mm, and more preferably 0.5 mm to 0.9 mm. In either case, the above-mentioned body wall thickness values ​​provide a balance between the size of the container (or the size of the chamber within the body wall, if the body wall thickness is increased) and the effectiveness of the therapeutic effect provided by the container.

[0058] The release characteristics of carbon monoxide released from the chamber through the body wall can be affected by the thickness of the body wall. In particular, a thinner body wall can result in a higher carbon monoxide release rate than a thicker body wall, at least when the same body wall material is used. In this regard, the desired carbon monoxide release rate can depend on the particular application of the container, such as the desired therapeutic effect, the total amount of carbon monoxide released, and / or the type of therapeutic treatment to be performed. That is, the above-mentioned body wall thickness value can be selected depending on the desired application.

[0059] As mentioned above, the partition / protrusion may be provided by a protrusion in the lid, but instead, such partition or protrusion may be provided on the bottom of the body (e.g., where the body is integrally formed but a lid is also used) and / or on the side (wall) of the body. In other words, the bottom and / or side body walls may have a thickness corresponding to the preferred protrusion length mentioned above.

[0060] The body preferably has a lower end that is positioned to face the lid in a direction from the lid toward the lower end when the lid and the body are connected to each other. The lower end preferably has an outwardly convex shape. By configuring the lower end to have an outwardly convex shape, it is possible to alleviate and / or reduce discomfort experienced by a subject when introducing the container into the subject's body, for example, when ingesting, e.g., swallowing, the container, or when introducing the container into the rectum.

[0061] Preferably, the lid has at least one flange, and the body has a body wall that at least partially defines the chamber. The flange is preferably configured to extend along at least a portion of the body wall when the lid and body are connected to each other. This configuration allows for a more secure and robust connection between the lid and body. Furthermore, the flange also functions as a guide when assembling the lid and body, making it easier to assemble the lid and body. The flange may be configured as a collar extending radially and / or axially relative to the longitudinal axis of the container.

[0062] The groove and the protrusion are preferably integrally provided on the lid. The groove is preferably at least partially defined by at least a portion of the flange and at least a portion of the protrusion, and is preferably configured so that at least a portion of the body, preferably at least a portion of the annular wall of the body, can be at least partially inserted between the protrusion and the flange to connect the body and the lid. This configuration makes it possible to more reliably and firmly connect the lid and the body. Furthermore, the flange and the protrusion also function as guides when assembling the lid and the body, making it easier to assemble the lid and the body.

[0063] The body and the lid are preferably connectable to one another by a material adhesive connection, preferably by an adhesive, preferably by a silicone-based adhesive, preferably by a biocompatible silicone, preferably by a medical grade silicone.

[0064] The overall size of the container is preferably a size corresponding to a standard pharmaceutical capsule size, preferably a size of a standard pharmaceutical capsule No. 5 to No. 000, or at least a size not exceeding this size.

[0065] The overall size of the container is preferably a size that corresponds to, or at least does not exceed, a standard pharmaceutical capsule size 2, 1, or 0.

[0066] The body and / or lid are preferably manufactured by a moulding process, preferably by a compression moulding process and / or an injection moulding process.

[0067] A second aspect of the present disclosure relates to a device for selectively releasing carbon monoxide, the device including a container according to any of the disclosed configurations. The device may further include at least one principal substance configured to selectively emit carbon monoxide. The principal substance may be stored within a chamber of the container. The principal substance may be configured to emit carbon monoxide when activated by at least one activating substance, and the emitted carbon monoxide may be released through at least a portion of a wall of the container.

[0068] The features and advantages detailed above with respect to the container apply equally to such devices.

[0069] Preferably, the entire device according to the second aspect of the present disclosure is configured as a capsule.

[0070] A third aspect of the present disclosure relates to a treatment system for treating biological cells, preferably living cells, preferably immune cells (such as CAR T cells and / or stem cells) and / or gametes with carbon monoxide. Such a treatment system may include at least one container configured to receive the biological cells, at least one source of carbon monoxide, and preferably at least one device for selectively releasing carbon monoxide according to the second aspect of the present disclosure. The container and the carbon monoxide source may be positioned relative to each other such that carbon monoxide provided by the carbon monoxide source can contact and treat the biological cells.

[0071] The configurations and advantages detailed above with respect to the vessel apply equally to such processing systems.

[0072] Such treatment systems may be configured to provide one or more effects on biological cells to at least partially preserve the biological cells.

[0073] Preferably, the biological cells are immune cells (such as CAR T cells and / or stem cells) and / or gametes. Preferably, the gametes include sperm or egg cells. Preferably, the sperm are contained in semen, glandular fluid, and / or a buffer system.

[0074] The biological cells may be human gametes and / or human immune cells. Alternatively or additionally, the gametes and / or immune cells may be of animal origin, preferably from cattle, horses, pigs, sheep, goats, camels, alpacas, dogs, and / or cats.

[0075] Such a container may be configured to preferably completely receive the biological cells and the source of carbon monoxide.

[0076] Such a container may include at least one compartment configured to receive a carbon monoxide-releasing device. In this case, the compartment is preferably housed and / or secured, preferably in a captive manner. This allows the treatment system to be pre-assembled, preferably before distribution to the application site, e.g., one or more laboratories, particularly by placing the carbon monoxide-releasing device in the compartment prior to use. This can facilitate handling and / or use of the treatment system and / or prevent the carbon monoxide-releasing device from being lost or separated from the container, e.g., during transport and / or handling.

[0077] The biological cells and the device for releasing carbon monoxide may be located within a single compartment within the container.

[0078] Alternatively, the biological cells and the device for releasing carbon monoxide may be located in separate compartments of the container. In this case, the compartments are preferably separated from one another by at least one wall and / or barrier permeable to carbon monoxide. The wall and / or barrier may have one or more openings configured to allow carbon monoxide to pass through and reach the biological cells. The compartment for receiving the carbon monoxide-releasing device is preferably located at and / or along the bottom of the container, particularly so that the biological cells are at least partially located above the compartment, particularly on the upper wall of the compartment. This allows for increased exposure and / or efficiency of exposure of the biological cells to carbon monoxide released from the device. Alternatively, the compartment may be located against and / or along the top and / or side of the container.

[0079] Such a container preferably includes a lid configured to form a seal with the container to seal the contents of the container from the environment, preferably airtight.

[0080] In a fourth aspect, the present disclosure relates to a set for treating, preferably biological cells, preferably living cells, with carbon monoxide, ex vivo or in vivo. The set includes at least one device for selectively releasing carbon monoxide according to the second aspect of the present disclosure. The set may further include at least one activating substance. Preferably, the activating substance is contained in, or at least provided in, an introduction device having a hollow needle configured to penetrate the device for selectively releasing carbon monoxide. More preferably, the hollow needle is configured to penetrate the lid of the device for selectively releasing carbon monoxide. The introduction device may be part of the set.

[0081] The configurations and advantages detailed above with respect to the containers apply equally to such sets.

[0082] A fifth aspect of the present disclosure relates to a method of treating a mammal, such as a human, with carbon monoxide in vivo. The method may include providing at least one device for selectively releasing carbon monoxide according to the second aspect of the present disclosure. The method may further include activating a principal substance with at least one activating agent by introducing the activating agent into a chamber of the device through a cover or body of the device. The method may further include introducing the at least one device into the mammal, preferably into one or more body cavities of the mammal, for example, by oral ingestion and / or rectal administration, and / or into the ear of an animal and / or human, for example (but not limited to) for the treatment of otitis media or other inflammatory conditions for which targeted release of CO may have therapeutic use, and / or into the vagina of an animal and / or human.

[0083] The features and advantages detailed above with respect to the container apply equally to such a method.

[0084] A sixth aspect of the present disclosure relates to carbon monoxide for treating biological cells (preferably living cells) and the interior and / or exterior of a mammalian body, preferably for use in treating gastrointestinal disorders, preferably released into and / or out of the mammalian body from a device for selectively releasing carbon monoxide, preferably from a device according to any of the embodiments described herein (see second aspect above).

[0085] The features and advantages detailed above with respect to the container apply equally to the sixth aspect.

[0086] A seventh aspect of the present disclosure relates to a method of treating biological cells (preferably living cells), more preferably ex vivo, with carbon monoxide. The method may include providing a treatment system according to the third aspect above. The method may include placing biological cells in the container. The method may include inducing release of carbon monoxide from the carbon monoxide source. Carbon monoxide provided by the carbon monoxide source may contact the biological cells.

[0087] The features and advantages detailed above apply equally to the seventh aspect.

[0088] Such a treatment system is preferably provided by disposing within the vessel at least one device configured to selectively release carbon monoxide, preferably at least one device according to any of the embodiments described herein with respect to the second aspect above.

[0089] Preferably, inducing the release of carbon monoxide includes activating the primary source of carbon monoxide with at least one activating substance.

[0090] Preferably, the activating substance is introduced into the chamber in which the principal substance is disposed, and the activating substance is preferably introduced by an introduction device, which may include a hollow needle configured to penetrate the wall of the chamber.

[0091] Such a treatment system preferably includes at least one first chamber and at least one second chamber. For example, biological cells can be placed in the first chamber. For example, at least a principal substance can be placed in the second chamber. The second chamber may be separated from the first chamber by at least one wall. At least a portion of the wall may be permeable to carbon monoxide emitted from the principal substance, thereby allowing the carbon monoxide emitted from the principal substance to be released from the second chamber into the first chamber.

[0092] Preferably, the release of carbon monoxide from such a device occurs at least within the digestive tract of a mammal, for example, for the treatment of digestive disorders, such as gastrointestinal disorders, for example, inflammatory bowel disease such as ulcerative colitis and / or Crohn's disease.

[0093] Such devices are preferably introduced into the mammalian body enterally, preferably by oral ingestion and / or rectal administration.

[0094] The following list of aspects provides preferred embodiments of the present disclosure.

[0095] 1. A container preferably adapted to be inserted into a human and / or animal body, at least one housing; at least one chamber defined within the housing; Including, the chamber is configured to receive and store at least one principal substance configured to selectively emit at least one gas, preferably at least carbon monoxide; the principal substance is configured to emit the gas after being activated by at least one activating substance; the chamber is enclosed by at least one wall of the housing; A container wherein at least a portion of the wall is at least partially permeable to the gas emitted by the principal substance, thereby allowing the container to at least partially release the gas emitted by the principal substance to the environment, and wherein the container is preferably configured such that the activating substance is introduced into the chamber from outside the housing, with the principal substance already disposed in the chamber.

[0096] 2. The container according to aspect 1, wherein the container includes only a single chamber.

[0097] 3. The container according to aspect 1 or 2, wherein the wall of the housing is impermeable to one or more of solids, liquids, toxic substances (preferably at least heavy metal atom-containing substances), and by-products (preferably at least toxic by-products) of the reaction between the principal substance and the activating substance.

[0098] 4. A container according to any one of aspects 1 to 3, wherein the container is configured to emit the gas (preferably carbon monoxide) in vivo and / or ex vivo.

[0099] 5. A container according to any of aspects 1 to 4, wherein said container is configured to be inserted into a human and / or animal body.

[0100] 6. A container according to any of aspects 1 to 5, wherein said container is adapted for oral and / or rectal administration to a human and / or animal body.

[0101] 7. A container according to any of aspects 1 to 6, wherein the container is configured to be ingested, preferably swallowed, by a human and / or animal.

[0102] 8. The container according to any of aspects 1 to 7, wherein at least one section of the housing is configured to be penetrated by an introduction device configured to introduce the activating substance into the chamber and activate the principal substance, and wherein the at least one section of the housing is configured to reseal, preferably substantially liquid-tight and / or air-tight, after the introduction device is withdrawn from the housing.

[0103] 9. A container according to any one of aspects 1 to 8, wherein the housing includes at least one main body and at least one lid, and the main body and the lid are configured to be connectable to each other, preferably substantially liquid-tight and / or air-tight.

[0104] 10. A container according to aspect 9, wherein the lid and / or the body includes a protrusion configured to protrude into the chamber when the lid and the body are connected to one another.

[0105] 11. A container according to aspect 10, wherein the protrusion is configured as a partition that can be penetrated by an introduction device configured to introduce the activating substance into the chamber and activate the principal substance, and the protrusion is configured to reseal, preferably substantially liquid-tight and / or air-tight, after the introduction device is withdrawn from the protrusion.

[0106] 12. A container according to aspect 10 or 11, wherein when the body and the lid are interconnected, the length of the protrusion that protrudes inwardly into the chamber and / or along the longitudinal axis of the lid and / or the body is at least 1 mm, more preferably at least 1.2 mm, more preferably at least 1.4 mm, more preferably at least 1.6 mm, more preferably at least 1.8 mm, more preferably at least 2 mm, more preferably at least 2.2 mm, more preferably at least 2.4 mm, more preferably at least 2.6 mm, more preferably at least 2.8 mm, more preferably at least 3 mm, more preferably at least 3.2 mm, more preferably at least 3.4 mm, more preferably at least 3.6 mm, more preferably at least 3.8 mm, more preferably at least 4 mm, more preferably at least 4.2 mm, more preferably at least 4.4 mm, more preferably at least 4.6 mm, more preferably at least 4.8 mm, more preferably at least 5 mm, more preferably at least 5.2 mm, more preferably at least 5.4 mm, more preferably at least 5.6 mm, more preferably at least 5.8 mm, and most preferably at least 6 mm.

[0107] 13. A container according to any of sides 9 to 12, wherein the body and / or the lid have a groove, the groove preferably being formed in a ring or semi-ring shape around the longitudinal axis of the body and / or the lid, and a portion of the body or the lid that does not have the groove is configured to be at least partially inserted into the groove to connect the body and the lid to each other.

[0108] 14. A container according to aspect 13, wherein when the lid and the body are connected to each other, the protrusion defines the groove at least partially, preferably along the outer periphery of the protrusion, and preferably the groove is formed at least partially coaxially around the protrusion.

[0109] 15. A container according to any one of aspects 9 to 14, wherein the body and / or the lid are made of silicone, preferably biocompatible silicone, preferably medical grade silicone.

[0110] 16. A container according to any one of sides 9 to 15, in which the entire lid is integrally formed.

[0111] 17. A container according to any one of sides 9 to 16, wherein the entire body is integrally formed.

[0112] 18. A container according to any one of aspects 9 to 17, wherein the lid and / or the body has a Shore A hardness of 20 Shore A to 80 Shore A, preferably 30 Shore A to 50 Shore A.

[0113] 19. A container according to any of aspects 9 to 18, wherein the body and / or the lid include at least one sealing element and / or friction-increasing element disposed along at least one connecting interface between the body and the lid, which increases the sealing action and / or frictional force between the body and the lid when the body and the lid are interconnected.

[0114] 20. A container according to side 19, wherein the sealing element and / or friction-increasing element includes at least one protrusion (e.g., rib) protruding from a boundary surface extending along the connecting interface of the lid and / or the body, and when the body and the lid are interconnected, the protrusion protrudes toward and engages with the lid or the body that does not have the protrusion.

[0115] 21. A container according to aspect 20, wherein the lid or the body includes the protrusion and the boundary surface extending along the joining interface of the lid or the body not having the protrusion is substantially smooth and / or continuous and / or free of protrusions and / or depressions.

[0116] 22. A container according to side 20 or 21, wherein the protrusion is formed at least partially along the circumference of the interface surface of the lid and / or body, preferably over multiple circumferences, preferably in a spiral and / or thread-like and / or serpentine and / or zigzag shape.

[0117] 23. The lid includes the protrusion; a distal end of the protrusion that protrudes inwardly from the body when the lid and the body are connected to each other has one or more rounded and / or chamfered edges; and / or A container relating to any of sides 9 to 22, in which at least a portion of the protrusion is tapered toward the distal end of the protrusion that penetrates deepest into the body when the lid and the body are connected to each other.

[0118] 24. A container according to any one of sides 20 to 23, wherein the protrusions and the projections are integrally and / or monolithically formed.

[0119] 25. A container according to any of aspects 1 to 24, wherein the greatest overall dimension of the container, preferably the length along the longitudinal axis of the container, is less than 25 mm, preferably less than 24 mm, more preferably less than 23 mm, more preferably less than 22 mm, more preferably less than 21 mm, more preferably less than 20 mm, more preferably less than 19 mm, more preferably less than 18 mm, more preferably less than 17 mm, more preferably less than 16 mm, more preferably less than 15 mm, more preferably less than 14 mm, more preferably less than 13 mm, more preferably less than 12 mm.

[0120] 26. The maximum dimension of the entire container in a direction perpendicular to the longitudinal axis of the body and / or the longitudinal axis of the lid, in a direction which extends substantially transverse to the front of the container, is less than 12 mm, more preferably less than 11 mm, more preferably less than 10 mm, more preferably less than 9 mm, more preferably less than 8 mm, more preferably less than 7 mm, more preferably less than 6 mm, more preferably less than 5 mm, more preferably less than 4 mm, of any of sides 1 to 25.

[0121] 27. The body has at least one body wall which at least partially defines the chamber, wherein the body wall is at least partially permeable to the gas which is released by the principal substance after the principal substance has been activated by the activating substance, and wherein the body wall has a thickness ranging from 0.4mm to 2.5mm, preferably from 0.5mm to 2.4mm, more preferably from 0.5mm to 2.3mm, more preferably from 0.5mm to 2.2mm, more preferably from 0.5mm to 2.1mm, more preferably from 0.5mm to 2mm, more preferably from 0.5mm to 1.9mm, more preferably from 0.5mm to 1.8mm, more preferably from 0.5mm to 1.7mm, more preferably from 0.5mm to 1.6mm, more preferably from 0.5mm to 1.5mm, more preferably from 0.5mm to 1.4mm, more preferably from 0.5mm to 1.3mm, more preferably from 0.5mm to 1.2mm, more preferably from 0.5mm to 1.1mm, more preferably from 0.5mm to 1mm, more preferably from 0.5mm to 0.9mm, a container relating to any of sides 9 to 26.

[0122] 28. A container according to any of sides 9 to 27, wherein the main body has a lower end, which is formed so as to protrude in a direction opposite to the lid when the lid and the main body are connected to each other, and the lower end has a convex external shape.

[0123] 29. A container according to any of sides 9 to 28, wherein the lid has at least one flange, the body has a body wall that at least partially defines the chamber, and the flange is configured to protrude along at least a portion of the body wall when the lid and the body are connected to each other.

[0124] 30. A container according to any of sides 9 to 29, wherein the groove and the protrusion are preferably integrally formed in the lid, preferably the groove is at least partially defined by at least a part of the flange and at least a part of the protrusion, and preferably a part of the body, preferably a part of the annular wall of the body, is configured to be at least partially insertable between the protrusion and the flange to connect the body and the lid to each other.

[0125] 31. A container according to any of aspects 9 to 30, wherein the body and the lid can be connected to each other by a material bond, preferably by a silicone adhesive, preferably a biocompatible silicone, preferably a medical grade silicone adhesive.

[0126] 32. A container according to any of aspects 1 to 31, wherein the overall size of the container corresponds to or at least does not exceed a standard pharmaceutical capsule size, preferably a standard pharmaceutical capsule size of 5,000.

[0127] 33. A container according to any of aspects 1 to 32, wherein the overall size of said container corresponds to or at least does not exceed a standard pharmaceutical capsule size 2, 1, or 0.

[0128] 34. A container according to any one of aspects 1 to 33, wherein the body and / or the lid are manufactured by a molding process, preferably a compression molding process and / or an injection molding process.

[0129] 35. An apparatus for selectively releasing at least one gas, preferably at least carbon monoxide, comprising: The container having any one of sides 1 to 34; at least one principal substance configured to selectively emit said gas; The device is configured such that the principal substance stored in the chamber of the container is activated by at least one activating substance and then emits the gas, thereby causing the gas to be released through at least a portion of the wall of the container.

[0130] 36. A treatment system for treating biological cells (preferably living cells, preferably immune cells and / or gametes) with at least one gas (preferably at least carbon monoxide), comprising: at least one container configured to receive biological cells; a source of at least one of said gases (preferably carbon monoxide), preferably a device for selectively releasing at least one gas (preferably at least carbon monoxide) according to at least one side 35; Including, A treatment system wherein the vessel and the source of gas are positioned relative to one another such that the gas supplied by the source of gas contacts and treats the biological cells.

[0131] 37. The treatment system according to aspect 36, wherein the container is configured to receive (preferably entirely) the biological cells and the source of the gas.

[0132] 38. A set for treating biological cells (preferably living cells) ex vivo or in vivo with at least one gas (preferably at least carbon monoxide), comprising: a device for selectively releasing at least one gas (preferably at least carbon monoxide) according to at least one side 35; At least one activator Including, Preferably, the activating substance is contained within an introduction device having a hollow needle configured to penetrate the device that selectively releases the gas, and more preferably, the hollow needle is configured to penetrate the lid of the device that selectively releases the gas.

[0133] 39. A method for treating a mammal, such as a human, in vivo with at least one gas, preferably at least carbon monoxide, comprising: providing a device for selectively releasing at least one gas (preferably at least carbon monoxide) according to at least one aspect 35; activating the principal substance with at least one activating substance, such as by introducing the activating substance into a chamber of the device through the lid or the body of the device; introducing said at least one device into said mammal, for example by oral ingestion and / or rectal administration, and / or into the ear of said mammal, and / or into the vagina of said mammal; The method includes:

[0134] 40. Carbon monoxide for use in treating biological cells (preferably living cells), inside and / or outside the body of a mammal, preferably for the treatment of a digestive disease (preferably an inflammatory disease of the digestive tract) or for digestive transplantation, wherein the carbon monoxide is released from a device according to aspect 35 into the body of the mammal and / or outside the body of the mammal.

[0135] 41. Carbon monoxide according to aspect 40, wherein the release of carbon monoxide from the device occurs within at least a portion of the mammal's digestive tract.

[0136] 42. Carbon monoxide according to aspect 40 or 41, wherein the device is introduced into the body of the mammal, for example into the intestine, preferably by ingestion and / or rectal administration, and / or into the ear of the mammal, and / or into the vagina of the mammal.

[0137] 43. Carbon monoxide according to aspect 40, 41, or 42, wherein the disease is one or more of diabetic gastroparesis, postoperative ileus, inflammatory bowel disease (e.g., ulcerative colitis and / or Crohn's disease), sepsis, gastritis, gastric ulcer, gastrointestinal ischemia-reperfusion injury, and non-alcoholic steatohepatitis.

[0138] 44. A method of treating biological cells (preferably living cells) with carbon monoxide (more preferably ex vivo), comprising: providing a processing system according to aspect 36; placing the biological cells in the container; Inducing the release of carbon monoxide from the carbon monoxide source, whereby the carbon monoxide provided by the carbon monoxide source contacts the living cells. The method includes:

[0139] 45. The method of aspect 44, wherein the treatment system is provided by disposing, within the vessel, at least one device configured to selectively release carbon monoxide (preferably at least one device according to aspect 35).

[0140] 46. ​​The method of aspect 44 or 45, wherein inducing the release of carbon monoxide comprises activating the primary substance of the carbon monoxide source with at least one activating substance.

[0141] 47. The method of aspect 46, wherein the activating substance is introduced into a chamber in which the principal substance is disposed, preferably wherein the activating substance is introduced via an introduction device, the introduction device including a hollow needle configured to penetrate the wall of the chamber.

[0142] 48. The method of any of aspects 44 to 47, wherein the treatment system includes at least one first chamber and at least one second chamber, wherein the biological cells are disposed in the first chamber and at least one principal substance is disposed in the second chamber, and preferably the second chamber is separated from the first chamber by at least one wall, and preferably at least a portion of the wall is permeable to carbon monoxide emitted from the principal substance, thereby allowing carbon monoxide emitted from the principal substance to be released from the second chamber into the first chamber.

[0143] Preferred embodiments of the present invention will be described in more detail below with reference to the drawings. The described embodiments do not limit the present invention. [Brief explanation of the drawings]

[0144] [Figure 1] FIG. 1 shows a schematic exploded view of a container according to one embodiment of the present disclosure. [Figure 2] FIG. 2 shows a further exploded view of the container shown in FIG. [Figure 3] FIG. 3 shows a further exploded view of the container shown in FIGS. [Figure 4] FIG. 4 is a diagram schematically illustrating the assembled state of the container of FIGS. [Figure 5] FIG. 5 shows a schematic side view of an apparatus for selectively releasing carbon monoxide, including the vessel shown in FIGS. [Figure 6] FIG. 6 is a schematic side view of a processing system according to one embodiment of the present disclosure. [Figure 7] FIG. 7 shows a schematic side view of a processing system according to a further embodiment of the present disclosure. [Figure 8] FIG. 8 is a diagram schematically showing a modified example of the lid of the container shown in FIGS. [Figure 9] Figure 9A shows the correlation between the volume of carbon monoxide released from the containers of Figures 1 to 8 and the mass of the main substance. Figure 9B shows the correlation between the volume of carbon monoxide released from the containers of Figures 1 to 8 and the volume of the activated substance. [Figure 10] FIG. 10 is a schematic diagram showing the results of an example of sperm treatment with carbon monoxide gas. [Figure 11] FIG. 11 is a further schematic diagram showing the results of an example of treating sperm with carbon monoxide gas. [Figure 12] FIG. 12 is a further schematic diagram showing the results of a further example of treatment of sperm with carbon monoxide gas. DETAILED DESCRIPTION OF THE INVENTION

[0145] 1-3 show exploded perspective views of a container 10 according to one embodiment of the present disclosure. FIG. 4 shows the container 10 of FIGS. 1-3 in an assembled state. The container 10 may be configured to be inserted into a human and / or animal body. Preferably, the container 10 is configured to be inserted, preferably manually, by ingestion, such as by swallowing the container 10, and / or rectally, and / or into the ear and / or vagina of a human and / or animal body, and / or into one or more body cavities of a human and / or animal body.

[0146] Container 10 may include at least one housing 12 and at least one chamber 14 defined within housing 12. Chamber 14 may be configured to receive and store at least one principal substance configured to selectively release carbon monoxide. In particular, the principal substance may be configured to emit carbon monoxide upon activation by at least one activating substance.

[0147] Chamber 14 may be surrounded by at least one wall 16 of housing 12. At least a portion of wall 16 may be at least partially permeable to carbon monoxide released by the principal substance, allowing container 10 to at least partially release carbon monoxide released by the principal substance to the environment, e.g., a treatment site within a subject's body. Container 10 may be configured such that an activating substance can be introduced into chamber 14 from outside housing 12, with the activating substance already disposed within chamber 14, by an introduction device, such as a syringe having a hollow needle configured to pierce a portion of housing 12.

[0148] The walls 16 of the housing 12 may be impermeable to one or more of solids, liquids, harmful substances (preferably at least heavy metal atom-containing substances), and by-products (preferably at least toxic by-products) resulting from the reaction of the principal substance with the activator substance.

[0149] 1-3, the container 10 may include only a single chamber 14. This allows the container 10 to be relatively compact. However, the container 10 may alternatively include multiple chambers 14.

[0150] The housing 12 may include at least one body 18 and at least one lid 20. The body 18 and lid 20 may be configured to be connected to one another, preferably in a substantially liquid-tight and / or air-tight manner, such that the chamber 14 is substantially sealed from the environment. The body 18 and lid 20 may be connected to one another, for example, by a friction-type connection, a form-locking connection, and / or a material adhesive connection. The material adhesive connection may be achieved by at least one adhesive, such as at least one silicone adhesive. The body 18 and / or lid 20 may be made of silicone. Preferably, the entire container 10 is made of a single material, for example, silicone.

[0151] As shown in Figures 1-3, the lid 20 may include a protrusion 22 configured to protrude into the chamber 14 when the lid 20 and the body 18 are coupled together.

[0152] The protrusion 22 may be configured as a septum that can be penetrated by an introduction device configured to introduce an active substance into the chamber 14 to activate the principal substance. A suitable introduction device may be a syringe having a hollow needle configured to penetrate the protrusion 22 and extend into the chamber 14 to inject the active substance into the chamber 14. Preferably, the protrusion 22 or septum is configured to reseal, preferably in a substantially liquid-tight and / or air-tight manner, after the introduction device is withdrawn from the protrusion 22.

[0153] Alternatively, or in addition, the body 18 may include a protrusion and / or a partition similar to or identical to the protrusion 22 shown in FIGS. 1-3. The protrusion and / or partition on the body may be configured to protrude into, or at least be disposed within, the chamber 14 when the lid 20 and the body 18 are coupled together. For example, the bottom wall of the body 18 (e.g., the end of the body 18 opposite the lid 20) and / or the side walls of the body 18 may provide a partition. To this end, the bottom wall and / or the side walls may have a thickness greater than the peripheral (e.g., cylindrical) portion of the wall 16. The bottom and / or side walls may have a thickness of at least 1.2 mm, preferably at least 2 mm, more preferably at least 3 mm, more preferably at least 4 mm, and even more preferably at least 5 mm.

[0154] As shown in Figures 1-3, the lid 20 may include a groove 24 (see Figures 2 and 3) that extends annularly or semi-annularly around the longitudinal axis 26 of the body 18 and / or the lid 20. As shown in Figures 1-3, the groove 24 may be formed at least partially coaxially around the protrusion 22.

[0155] The lid 20 may include at least one flange 25 that may be configured to extend along at least a portion of the wall 16 when the lid 20 and the body 18 are coupled together (see FIG. 4).

[0156] 1 to 3, the groove 24 and the protrusion 22 may be preferably integrally provided on the lid 22. As shown in FIGS. 1 to 3, the groove 24 may be at least partially defined by at least a portion of the flange 25 and at least a portion of the protrusion 22, and may preferably be defined along the outer periphery 28 of the protrusion 22. A portion of the wall 16 may be at least partially inserted between the protrusion 22 and the flange 25, i.e., into the groove 24, and may assist in connecting the main body 18 and the lid 20.

[0157] Alternatively, or in addition, as shown in Figures 1-3, body 18 may include a groove similar to or identical to groove 24 provided in lid 20, as shown in Figures 1-3. A portion of lid 20 may be configured to be at least partially inserted into groove provided in body 18 to facilitate coupling of body 18 and lid 20 to one another.

[0158] Figure 5 shows an apparatus 40 for selectively releasing carbon monoxide, including the container 10 shown in Figures 1-4 and a carbon monoxide-releasing primary substance 42. As shown in Figure 5, the primary substance 42 may be stored within the chamber 14 of the container 10 and may be configured to release carbon monoxide after activation by at least one activating material 44, such that the released carbon monoxide is released through at least a portion of the wall 16 of the container 10.

[0159] The depiction of the primary substance 42 and the activator substance 44 in Figure 5 is merely exemplary and schematic. The primary substance 42 is preferably a powder and / or granules. Alternatively, the primary substance 42 may be a liquid and / or gas. Similarly, the activator substance 44 may be a powder, granules, liquid, and / or gas.

[0160] The activating substance 44 may be introduced into the chamber 14 of the container 10 by an introduction device 46. As shown in Figure 5, the introduction device 46 may be configured as a syringe with a hollow needle 48 configured to penetrate the lid 20, and more specifically, the protrusion 22, into the chamber 14 to introduce the activating substance 44 and activate the principal substance 42. The lid 20 may be configured to reseal after the introduction device 46 is withdrawn from the lid 20, preferably in a substantially liquid-tight and / or air-tight manner.

[0161] As noted above, container 10, and more specifically device 40, shown in Figures 1-5 may be configured for treating an animal and / or human body in vivo by releasing carbon monoxide in vivo. Preferably, container 10, and more specifically device 40, is configured for insertion, preferably manually, by ingestion, such as by swallowing container 10, and / or rectally, and / or into the ear and / or vagina of a human and / or animal body, and / or into one or more body cavities of a human and / or animal body. This allows for treatment of living cells in a human and / or animal body with carbon monoxide released from container 10, and more specifically device 40.

[0162] Alternatively, however, the container 10, and more particularly the device 40, may be configured to treat biological cells, preferably living cells, extracorporeally by releasing carbon monoxide extracorporeally. Such a treatment system 60 is shown in FIG. 6. The treatment system 60 is configured to treat biological cells 62, preferably living cells, preferably immune cells and / or gametes, with carbon monoxide. The treatment system 60 may be configured to impart one or more effects to the biological cells 62, such as at least partially preserving the biological cells 62. The treatment system 60 may include at least one container 64 configured to receive the biological cells 62 and at least one source of carbon monoxide, and in the configuration shown in FIG. 6, is embodied by the device 40 shown in FIG. 5. The container 64 and the carbon monoxide source device 40 may be positioned relative to one another such that the biological cells 62 are contacted with the carbon monoxide provided by the device 40, thereby treating the biological cells 62. Container 64 may preferably include a lid 66 configured to form a seal with container 64 to seal, preferably airtight, the contents of container 64 from the environment. Alternatively, or in addition, treatment system 60 may be configured to treat an animal and / or human body extracorporeally, for example, by applying carbon monoxide to an external surface of the animal and / or human body, such as the skin, which may also be considered treatment of living cells within the meaning of the present disclosure.

[0163] FIG. 7 shows a schematic diagram of an alternative treatment system 80 for treating biological cells, preferably live cells, ex vivo by releasing carbon monoxide ex vivo. The treatment system 80 may include at least one container 82 configured to receive biological cells 62 and at least one source of carbon monoxide, embodied by, by way of example and not limitation, the device 40 shown in FIG. 5 in the configuration shown in FIG. 7 . The container 82 and the carbon monoxide source device 40 may be positioned relative to one another such that carbon monoxide provided by the device 40 contacts the biological cells 62 for treatment. The container 82 may include at least one compartment 84 configured to receive, preferably encase and / or secure, preferably constrainingly, the device 40. Alternatively, or in addition to the compartment 84, the container 82 may include one or more securing means, e.g., one or more clips, configured to secure the carbon monoxide-releasing device 40 within the container 82, preferably captively and / or substantially immovably. As a further alternative, device 40 may be omitted, and principal substance 42 may be placed directly in at least one chamber, e.g., compartment 84, defined within container 82. For example, compartment 84 may itself form container 10, and, for example, the walls of compartment 84 may form, or at least a portion of, housing 12 of container 10. This may allow processing system 80 to be pre-assembled by placing device 40 within compartment 84, particularly prior to use, and preferably before distribution to the application site, e.g., one or more laboratories. This may facilitate handling and / or use of processing system 80.

[0164] Preferably, compartment 84 is disposed at and / or along the bottom of vessel 82, particularly such that biological cells 62 are disposed at least partially above compartment 84, particularly on an upper wall 86 of compartment 84, when treatment system 80 is in an operational state. Upper wall 86 of compartment 84 may be permeable to carbon monoxide emitted from device 40. This arrangement may increase exposure of biological cells 62 to carbon monoxide emitted from device 40. Upper wall 86 may include one or more openings 88 configured to allow carbon monoxide to pass through the upper wall and reach biological cells 62. Alternatively, compartment 84 may be disposed on and / or along an upper surface and / or at least one side surface of vessel 82.

[0165] Container 82 is preferably configured to receive multiple devices 40 in addition to (and apart from) the compartments configured to receive biological cells 62, e.g., by including multiple sources of carbon monoxide, e.g., multiple devices 40, e.g., multiple compartments 84. Each compartment 84 is configured to receive at least one source of carbon monoxide, e.g., device 40.

[0166] Container 82 may preferably include a lid 90 configured to form a seal with container 82, preferably in an airtight manner, that seals the contents of container 82 from the environment. Alternatively or additionally, treatment system 80 may be configured to treat an animal and / or human body extracorporeally, for example, by applying carbon monoxide to an external surface of the animal and / or human body, such as the skin, which may also be considered treatment of living cells within the meaning of the present disclosure.

[0167] Any type of trigger mechanism, such as device 40, for releasing carbon monoxide from a carbon monoxide source may be provided, particularly when the carbon monoxide source is located within container 82, more specifically within compartment 84, and / or when cell 62 is located within container 82.

[0168] Figure 8 schematically illustrates a further embodiment of the lid 20 for the container 10 shown in Figures 1-6. The lid 20 shown in Figure 8 includes at least one protrusion 70 (e.g., a rib) extending from an interface surface 72 of the lid 20. The interface surface 72 may protrude along a connection interface between the lid 20 and the body 18. For example, as shown in Figures 4-6, the protrusion 70 may extend toward the body 18, for example, along an inner surface of the body 18, and engage with the body 18 when the body 18 and the lid 20 are connected to one another.

[0169] The protrusions 70 may be at least partially formed in a spiral and / or thread-like and / or serpentine and / or zigzag shape, preferably along the circumference, preferably along the entire circumference, preferably over multiple revolutions, of the interface surface 72 of the lid 20, as exemplarily shown in Figure 8. This may provide a relatively large surface over which the protrusions 70 may provide a higher sealing effect and / or higher friction and / or higher connection force.

[0170] The boundary surface of body 18 extending along the mating interface between lid 20 and body 18 is preferably substantially smooth and / or continuous and / or devoid of protrusions and / or depressions. Alternatively, body 18 may have one or more protrusions and / or one or more recesses, such as one or more recesses configured to receive protrusion 70.

[0171] Alternatively, or in addition, body 18 may include at least one protrusion configured similarly or identically to protrusion 70 .

[0172] The distal end 76 of the projection 22, which penetrates furthest into the body 18 when the lid 20 and body 18 are connected together, may have one or more rounded and / or curved edges 94. Alternatively, or additionally, the projection 22 may be tapered along at least a portion of the projection 22 toward the distal end 76 of the projection 22. This may facilitate manufacturing of the lid 20, for example, by making it easier to remove the molded lid from a mold compared to a cylindrical shape of the lid 20 / projection 22, or by reducing the risk of damage during removal if the lid 20 is manufactured by a molding process. Additionally, this may prevent, or at least reduce, spillage of a substance, such as an active substance, after removal of an introducer device, e.g., a hollow needle, such as the introducer device 46 shown in FIG. 5 . For example, compared to configuring the protrusion 22 to have one or more sharp edges at the distal end 76 of the protrusion 22, leaving space or additional space at the edge of the distal end 76 of the protrusion 22 prevents or at least reduces spillage of a substance, such as an active substance, after removing the introduction device of FIG. 5, which shows a hollow needle as the introduction device 46 for introducing a substance into the container 10.

[0173] The lid 20 shown in FIG. 8 may include the groove 24 shown in FIGS.

[0174] 9A shows the correlation between the amount / volume (mL) of carbon monoxide (CO) released from the container 10 and the mass (mg) of the primary substance 42 ("Mo-CORM") after the activated material 44 and the primary substance 42 were combined in the chamber 14 of the container 10. The amounts of carbon monoxide (CO) released were 1.30 mL, 0.95 mL, 0.44 mL, and 0.23 mL, which corresponded to 15 mg, 10 mg, 5 mg, and 2.5 mg of the primary substance 42 ("Mo-CORM"), respectively.

[0175] 9B shows the correlation between the amount of carbon monoxide (CO) released per volume (mL) from the container 10 and the volume (μL) of activating material 44 (“FeCl”) after the activating material 44 and the principal material 42 were combined in the chamber 14 of the container 10. The amount of carbon monoxide (CO) released ranged from 0.95 mL to 0.34 mL, corresponding to 50 μL to 10 μL of activating material 44 (“FeCl”). [Example]

[0176] Example of treatment of sperm with carbon monoxide:

[0177] The following examples demonstrate the improvement of sperm characteristics (oxidation-reduction potential, DNA fragmentation and / or sperm motility, especially forward motility) upon exposure to carbon monoxide gas.

[0178] In particular, Example 1 relates to the treatment of sperm with carbon monoxide released from a treatment system comprising a capsule with two compartments, as described in more detail below. The results of Example 1 are shown in Figures 10 and 11.

[0179] Example 2 relates to the treatment of sperm using carbon monoxide released from a therapeutic system including a container, e.g., a capsule, having a single chamber or compartment defined within a housing, configured similarly or identically to any of the embodiments described herein with respect to the first aspect of the present disclosure, e.g., according to any of the embodiments described herein. The results of Example 2 are shown in Figure 12.

[0180] The results of both examples below show that for various treatment systems, here the different treatment systems used in Examples 1 and 2, sperm properties (oxidation-reduction potential, DNA fragmentation and / or sperm motility, particularly forward motility) are improved when sperm are exposed to carbon monoxide gas.

[0181] Example 1: method In this example, a CORS capsule measuring 18–22 mm in length and 6–8 mm in diameter was used to store 15 mg of CO-releasing molecule (Mo-CORM) and 150 μL of 583.3 mg / mL FeCl3 aqueous solution in a standard 15 mL Falcon tube. The CORS, Mo-CORM, and FeCl3 were prepared as follows:

[0182] The capsules, CORS, and FeCl3 were prepared as follows.

[0183] Preparation of capsules The capsule was fabricated using additive manufacturing. Two compartments, separated by a partition, were 3D printed within one compartment. The compartment containing the aqueous solution of ferric chloride was printed using a drop-on-demand (PolyJet modeling process) method using an Objet Eden 350 printer (96 nozzles, 40 μM droplet size, resolution: X = 600 dpi, Y = 600 dpi; Stratasys, Deden Prairie, MN, USA). The photopolymerizable resin VeroBlackPlus (Stratasys, Deden Prairie, MN, USA) was used, and the structure was cured by UV polymerization at 365 nm using a UV lamp. A water-soluble support structure (SUP705; Stratasys, Deden Prairie, MN, USA) was used and removed by washing (potable water; isopropanol) after the printing process.

[0184] The compartment housing the CORM was fabricated by laser sintering (LS) using polyamide PA2200 (EOS GmbH, Krailing, Germany). Processing was performed using a Formiga P110 3D printer (EOS GmbH, Krailing, Germany) with a CO2 laser (wavelength 10.6 μm, layer thickness 0.1 mm). The silicone membrane (SIK8649) surrounding the two compartments was obtained from RAUMEDIC AG (Helmbreckt, Germany). CAD software (VISI 2019 & 2020, Vero UK, Cheltenham, UK) was used for the production-ready design.

[0185] Preparation of CORM The synthesis of trisodium tricarbonyl-[tris(isocyanethylacetato)]molybdenum CORM (NaMo(CO)(CNCHCO)), Mo-CORM, was carried out with some modifications according to a previous protocol (Achatz, D et al. Zeitschrift für anorganische und allgemeine Chemie 2005, 631 (12), 2339-2346). Briefly, under dry and inert conditions, 2.64 g (10.0 mmol) of molybdenum hexacarbonyl was dissolved in 35 mL of anhydrous acetonitrile (99.8%; 90 °C; 22 h) to form the intermediate acetonitrile complex. Subsequently, 4.7 mL of ethyl isocyanate (3.5 equiv, 35 mmol, 4.86 g) was added at 55 °C to displace the acetonitrile and form the complex. The ester was then hydrolyzed with 20 mL of NaOH (16 equiv.) in tetrahydrofuran at room temperature. The free acid was formed by adding aqueous hydrochloric acid. Finally, the trisodium salt was obtained by adding NaOH in ethanol (5 mmol NaOH / 1 mmol Mo-CORM). All reagents were purchased from Sigma Aldrich (Schnelldorf, Germany) and used without further purification.

[0186] Preparation of FeCl3 aqueous solution FeCl3·6H2O was purchased from Sigma Aldrich (Schnelldorf, Germany) and prepared in deionized water to a concentration of 583.3 mg / mL to prepare an aqueous solution.

[0187] sperm analysis Semen / sperm volumes of 1.46 mL to 5.7 mL were collected in 100 mL sample vials from healthy men aged 25 to 50 years. The samples were then divided into two equal fractions, each incubated in the presence (wCO) and absence (woCO). To estimate the sperm concentration in the samples and establish an initial baseline for each sample, a routine sperm analysis was performed before transferring the samples to gas-tight 15 mL Falcon tubes. In a sealed container, the samples were incubated at 25–35°C for 90 minutes with and without an activated CORS system. Liquefaction occurred naturally during this incubation period. The samples were then analyzed for total motility, forward motility, resting redox potential, and DNA fragmentation.

[0188] Sample total motility, forward motility, and concentration were assessed using the CEROS II (Hamilton Thorne, Beverly, MA) computer-assisted sperm analysis (CASA) system. Six microliters of sperm sample diluted 1:1 with Multipurpose Handling Medium-Complete (MHM-C, Fujifilm / Irvine Scientific, Santa Ana, CA) were measured to assess concentration and motility (total motility and forward motility).

[0189] Static oxidation-reduction potential (sORP) levels were measured using the Male Infertility Oxidative System (MiOXSYS, Englewood, CO). A 30 μL sperm sample was transferred to the sample application port of the MiOXSYS sensor and analyzed for 2 minutes.

[0190] A count-based microscopy assay was performed to measure DNA fragmentation. The sperm samples were diluted to a maximum of 20xE6 sperm per mL using MHM-C (Multipurpose Handling Medium-Complete, Fujifilm, Irvine Scientific) and analyzed using the Halo Sperm G2 kit (Halotech, Madrid, Spain). Prepared agarose was mixed with the sample according to the manufacturer's instructions. 8 μL of the resulting mixture was placed on a microscope slide, removed from the heat source, and allowed to cool for 5 minutes. The prepared samples were treated with denaturant for 7 minutes, lysis solution for 20 minutes, distilled water for 5 minutes, 70% ethanol for 2 minutes, 100% ethanol for 2 minutes, eosin stain for 7–10 minutes, and thiazine stain for 7–10 minutes according to the manufacturer's instructions. At least 300 sperm were then analyzed using bright-field microscopy.

[0191] result 1. (Static) Oxidation-Reduction Potential (sORP) Level The "relative redox potential" data points shown in Figure 2 are derived for the following ratio per subject: sORP wCO (Subject n) / sORP woCO (Subject n), where sORP woCO (Subject n) is set to 1. The sORP data, which is an indicator of oxidative stress (e.g., due to ROS) in sperm (spermatozoa), tend to be lower in samples exposed to CO (wCO) than in samples not exposed to CO (woCO). This is evident from the location of the data points; four of these values ​​are located below the dashed reference line for woCO = 1. Therefore, without being bound by theory, these results suggest that the levels of reactive oxygen species (ROS) in sperm are reduced compared to sperm exposed to CO and those not exposed to CO (Figure 10).

[0192] 2. DNA Fragmentation The "relative DNA fragmentation" data points shown in Figure 11 are values ​​derived for each subject, where the ratio for each subject is: DNA fragmentation wCOSubject n / DNA fragmentation woCOSubject n, where DNA fragmentation woCOSubject n is set to 1. As is clear from Figure 11, samples exposed to CO have reduced DNA fragmentation compared to samples not exposed to CO. This is evident from the location of the data points; all of these data points are located below the dashed reference line for woCO=1.

[0193] In summary, these results indicate that sperm exposure to CO leads to desirable / improved gamete characteristics, such as improved progressive motility, as well as reduced oxidative stress / redox potential / reactive oxygen levels (Figure 10) and DNA fragmentation (Figure 11).

[0194] Thus, the present inventors have surprisingly found that contacting gametes, such as sperm, with carbon monoxide (CO) preserves the sperm. Incubating gametes with CO has shown that these sperm exhibit desirable properties (compared to sperm that have not been exposed to CO), making them particularly useful for ART.

[0195] Example 2: Example 2 demonstrates the improvement of sperm viability parameters (i.e., oxidation-reduction potential) upon carbon monoxide (CO) incubation for different incubation times. Sperm collected from the same subject were either treated with carbon monoxide or left untreated as "untreated" controls. Carbon monoxide treatment was performed for various treatment times (60, 90, 180, or 270 minutes).

[0196] method In this example, sperm samples were processed in one, three, or six containers / capsules, each loaded with 5 mg of Mo-CORM (equivalent to 0.44 mL of CO) and processed for 60 to 270 minutes, or left unprocessed for the same time periods.

[0197] The container and Mo-CORM were prepared using the materials described below: FeCl3 was used to activate the Mo-CORM.

[0198] material All chemicals required for the synthesis and activation of Mo-CORM (molybdenum hexacarbonyl, ethyl isocyanoacetate, acetonitrile, anhydrous tetrahydrofuran, sodium hydroxide (pa), hydrochloric acid (pa), absolute ethanol, ferric chloride 6H2O, and nitric acid (65%, pa)) were purchased from Sigma-Aldrich Chemie GmbH (Schnelldorf, Germany). Multipurpose Handling Medium (MHM) for sperm dilution was purchased from Irvine Scientific (Santa Ana, CA). 270 ppm CO2 calibration gas was purchased from Linde AG (Munich, Germany). Polyamide (PA2200) was obtained from EOS GmbH (Kreiling, Germany), and Duroplast photopolymerizable resin MED610 + VeroBlackPlus (RGD875) was obtained from Stratasys Ltd. (Rehovot, Israel). Silicone R 6.65 × 0.4 mm was purchased from RAUMEDIC AG (Helmbrechts, Germany). SF33 2K silicone (mixing ratio 1:1, viscosity before mixing: 7000–8000 cP, hardness after mixing: 33 ShA, density 1.11 g / cc, break point: 4.7 N / mm) was used. 2 ) and SF45 2K silicone (mixing ratio 1:1, viscosity before mixing: 8500 cP (23°C), hardness after mixing: 45 ShA, density: 1.12 g / cc, breaking point: 3.5 ± 0.5 N / mm 2) were purchased from Siliconfabrik (Ahrensburg, Germany). All molds were made from polytetrafluoroethylene (Vink, Germany) using a CNC machine type neo (Datron AG, Germany). Loctite SI 5248, Loctite 4902, Loctite HY 4011, and Loctite SF7701 were purchased from Henkel (Düsseldorf, Germany). Sterican (登録番号) Needles 0.9 x 40 mm and Omnican (registration number) U100 insulin needles 0.3 x 8 mm were purchased from B. Braun (Melsung, Germany). All other reagents were purchased from Sigma Aldrich Chemie GmbH and were of at least pharmaceutical grade unless otherwise stated.

[0199] Container manufacturing All parts were molded using custom-designed Teflon® molds. After pouring the material into the mold, a vacuum was applied for five minutes to remove any air bubbles. The material was then poured again to the edges and vacuumed for another five minutes. After this time, a corresponding mold was pressed into the silicone and secured with a clamp. After drying at 40°C for 24 hours, the mold was separated from the mold with compressed air. The housing and lid were molded separately. The chamber was filled with Mo-CORM. Then, for some versions, an adhesive primer was applied to the lid and top edge of the housing. The adhesive was then applied, and both parts were pressed together. A circular motion between the two parts ensured even distribution of the adhesive.

[0200] Synthesis of Mo-CORM The CORM trisodium tricarbonyl-[tris(isocyanethylacetate)]molybdenum (Na3Mo(CO)3(CNCH2CO2)3, Mo-CORM) was synthesized as previously described (Reilaender, ACS Biomaterials Science & Engineering, 2022). Briefly, molybdenum hexacarbonyl was stirred in anhydrous acetonitrile, and the acetonitrile was exchanged with three ligands. These ligands were then exchanged with the EICA ligand. Purification and cation exchange afforded Mo-CORM as a white solid.

[0201] Sperm preparation Human sperm were collected from healthy volunteers in 100 mL containers. The glandular fluid was then aliquoted and placed into 15 mL gas-impermeable Falcon tubes. One sample was exposed to one or more activation containers / capsules. Both samples were then analyzed for static oxidation-reduction potential (sORP) using the MiOXSYS after a set time period.

[0202] Static Oxidation-Reduction Potential (sORP) Analysis Static oxidation-reduction potential (sORP) levels were measured using an Oxidative Male Infertility Measurement System (MiOXSYS, Englewood, Colorado) according to the manufacturer's instructions. A 30 μL sample was transferred to the sample application port of the MiOXSYS sensor and analyzed for 2 minutes. Results were expressed in millivolts (mV) and expressed as semen sperm concentration (mV / 10 6 Normalized to sperm / mL.

[0203] result FIG. 12 clearly demonstrates the beneficial effect of carbon monoxide on the viability parameters of gametes, i.e., the redox potential of the treated gametes. This is particularly evident in the gamete samples from subjects 38_7 and 42_1, which were exposed to carbon monoxide for 60 and 90 minutes, respectively. While both subjects' samples exhibit high redox potentials when untreated (black bars), a significant decrease in redox potential is observed in each sample when treated with carbon monoxide (white bars). While measuring redox potential is an indicator of oxidative stress in gametes, a high redox potential (such as in the untreated gamete samples from subjects 38_7 and 42_1) can adversely affect gamete viability and quality. Thus, the present inventors surprisingly discovered that exposure of gametes, such as sperm, to carbon monoxide (CO) improves gamete viability parameters and quality.

Claims

1. A container (10) configured to be inserted into a human and / or animal body, comprising: At least one housing (12); At least one chamber (14) defined within said housing (12); Including, the chamber (14) is configured to receive and store at least one principal substance (42) configured to selectively emit carbon monoxide; the primary material (42) is configured to emit carbon monoxide when activated by at least one activating material (44); The chamber (14) is surrounded by at least one wall (16) of the housing (12); at least a portion of the wall (16) is at least partially permeable to carbon monoxide emitted by the primary substance (42), thereby enabling the container (10) to at least partially release carbon monoxide emitted by the primary substance (42) to the environment; A container (10) configured such that the activating substance (44) can be introduced into the chamber (14) from outside the housing (12) with the principal substance (42) already disposed within the chamber (14).

2. The container (10) of claim 1, wherein the container (10) includes only a single chamber (14).

3. 3. The container (10) of claim 1 or 2, wherein the container (10) is configured to release carbon monoxide in vivo and / or ex vivo.

4. 4. The container (10) according to any one of claims 1 to 3, wherein the container (10) is adapted to be administered orally and / or rectally and / or vaginally and / or aurally to a human and / or animal body.

5. Container (10) according to any one of claims 1 to 4, wherein the container (10) is adapted to be swallowed by a human and / or an animal.

6. The container (10) according to any one of claims 1 to 5, wherein the housing (12) comprises at least one body (18) and at least one lid (20), the body (18) and the lid (20) being configured to be connectable to each other, preferably configured to be connectable in a substantially liquid-tight manner.

7. 7. The container (10) of claim 1, wherein the lid (20) and / or the body (18) includes a protrusion (22), and the protrusion (22) is configured to protrude into the chamber (14) when the lid (20) and the body (18) are connected to each other.

8. The protrusion (22) is configured as a partition wall, the septum is penetrable by an introduction device (46) configured to introduce the activating substance (44) into the chamber (14) to activate the principal substance (42); 8. The container (10) of claim 7, wherein the protrusion (22) is configured to reseal, preferably substantially liquid-tight, after the introduction device (46) is withdrawn from the protrusion (22).

9. the body (18) and / or the lid (20) includes at least one protrusion (70) protruding from a boundary surface (72) of the lid (20) and / or the body (18), the boundary surface (72) being a surface extending along a joining interface between the lid (20) and the body (18); When the body (18) and the lid (20) are connected to each other, the protrusion (70) is configured to protrude toward and engage with the other of the lid (20) and the body (18); Preferably, the container (10) according to any one of claims 6 to 8, wherein the lid (20) or the body (18) includes the protrusion (70) and the interface of the other of the lid (20) and the body (18), extending along the joining interface between the lid (20) and the body (18), is substantially smooth and / or continuous and / or free of protrusions and / or depressions.

10. The overall size of the container (10) corresponds to or at least does not exceed the standard pharmaceutical capsule size, and preferably the standard pharmaceutical capsule size is 5,000. and / or The container (10) of any one of claims 1 to 9, wherein the overall size of the container (10) corresponds to, or at least does not exceed, 2, 1, or 0 standard pharmaceutical capsule sizes.

11. A device (40) for selectively releasing carbon monoxide, comprising: A container (10) according to any one of claims 1 to 10; at least one primary substance (42) configured to selectively emit carbon monoxide; The device (40) is configured such that the principal substance (42) is stored within the chamber (14) of the container (10) and, upon activation by at least one activating substance (44), emits carbon monoxide, and the emitted carbon monoxide is released through at least a portion of the wall (16) of the container (10).

12. A treatment system (60; 80) for treating biological cells (62), preferably living cells, preferably gametes and / or immune cells, with carbon monoxide, comprising: at least one container (64; 82) configured to receive said biological cells (62); at least one source (40) of carbon monoxide, preferably at least one device (40) for selectively releasing carbon monoxide according to claim 11; Including, A treatment system (60; 80) in which the container (64; 82) and the carbon monoxide source (40) are arranged relative to one another so that the biological cells (62) come into contact with carbon monoxide supplied by the carbon monoxide source (40) and the biological cells (62) can be treated with carbon monoxide.

13. A set for treating biological cells (62), preferably living cells, with carbon monoxide ex vivo or in vivo, comprising: A device (40) for selectively releasing carbon monoxide according to at least one claim 11; at least one activator (44); Including, Preferably, the activating substance (44) is contained within an introduction device (46) having a hollow needle (48), the hollow needle (48) configured to penetrate the device (40) for selectively releasing carbon monoxide, and more preferably, the hollow needle (48) configured to penetrate the lid (20) of the device (40) for selectively releasing carbon monoxide.

14. 1. A method of preparing a carbon monoxide delivery device, comprising:

12. A device for selectively releasing carbon monoxide according to at least one claim 11, said device being adapted to be inserted into one or more body cavities of a human and / or animal body, preferably by oral ingestion and / or rectal insertion; activating the principal substance (42) with at least one activating substance (44) by introducing the activating substance (44) into the chamber (14) of the device (40) through the lid (20) or body (18) of the device (40); The method includes:

15. 1. A method of treating a mammal, such as a human, with carbon monoxide in vivo, comprising:

12. A device for selectively releasing carbon monoxide according to at least one claim 11, activating the principal substance (42) with at least one activating substance (44), such as by introducing the activating substance (44) into the chamber (14) of the device (40) through the lid (20) or body (18) of the device (40); Introducing the at least one device (40) into one or more body cavities of the mammal, such as by oral ingestion and / or rectal insertion. The method includes:

16. Carbon monoxide for use in treating biological cells, preferably living cells, inside and / or outside the body of a mammal, preferably for use in treating a digestive disorder, said carbon monoxide being released inside and / or outside the body of said mammal from a device according to claim 11.

17. 1. A method of treating biological cells (62), preferably living cells, more preferably ex vivo, with carbon monoxide, comprising: A processing system according to claim 12 is provided, placing said biological cells (62) in said container (64; 82); Inducing the release of carbon monoxide from the carbon monoxide source and contacting the carbon monoxide provided by the carbon monoxide source with the biological cells (62). The method includes:

18. 18. The method of claim 17, wherein the treatment system is provided by placing in the vessel (64; 82) at least one device (40) configured to selectively release carbon monoxide, preferably at least one device (40) according to claim 11.

19. 19. The method of claim 17 or 18, wherein inducing the release of carbon monoxide comprises activating the carbon monoxide source principal substance (42) with at least one activating substance (44).

20. The activating substance (44) is introduced into the chamber (14) containing the principal substance (42), and preferably the activating substance (44) is introduced via an introducing device (46); 20. The method of claim 19, wherein the introducer (46) comprises a hollow needle (48) configured to penetrate the wall of the chamber.

21. 21. The method of claim 17, wherein the treatment system comprises at least one first chamber and at least one second chamber, the biological cells (62) being disposed in the first chamber, and at least one carbon monoxide source principal substance (42) being disposed in the second chamber, the second chamber being separated from the first chamber by at least one wall (16; 86), at least a portion of which is permeable to carbon monoxide emitted by the principal substance (42) such that carbon monoxide emitted by the principal substance (42) is released from the second chamber into the first chamber.

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