An anti-aggregate device

Field of the invention

The present invention relates to an anti-aggregate device, and its use in the treatment, prevention and/or alleviation of symptoms of a disease, in particular wherein infusion of a cell suspension is necessary, as well as in nutrition.

Background

Cell therapies, in particular tumour infiltrating lymphocytes, CAR-T cells or stem cells, have the potential of revolutionizing the state of the art medical practice, in particular in the fields of oncology and haematology. In several applications, cell therapies involve infusing a subject (or a patient) with a cell suspension of particular cells, as recited hereinabove. As the production of such cell suspensions continues to pose significant challenges and is thus expensive, any loses of cells due to aggregation, for example while in the infusion bag, translate to significant cost and burden and should be avoided. One possibility to avoid aggregation is to add chemical agents into the cell suspension, which however limits the applicability of the cell suspension for infusing patients and is preferably avoided by clinical practitioners. As known to the skilled clinician, aggregation of cells in the cell suspension may be mostly happening during the infusion, to date, there appears to be no available solution that allows for mixing the cells in the infusion bag in the course of infusion, other than manual intervention.

The patent EP 728,491 discloses a certain blood transferring device equipped with ion concentration adjusting room.

The patent US 9,044,547 discloses a certain device and a method for preparing adipose tissue for transplantation from lobular fat extracted e.g. by liposuction.

The document CN 20,831 ,96411 discloses certain infusion set for on-the-spot use, comprising a ball pump.

Document US 2006/100578 discloses certain medication delivery apparatus and methods for intravenous infusions.

Document WO 1994/009847 discloses certain infusion pump, bag and method of use thereof.

Document US 2019/075830 discloses certain devices and methods for the preparation of a nutritional formula.

Summary of the invention

It was an objective technical problem of the present invention to provide the means for preventing aggregation of cells in the cell suspension.

The objective technical problem is solved by the embodiments described herein and as characterized in the claims.

The present inventors have surprisingly found that by mixing the cell suspension in the infusion bag during the infusion process, aggregation of the cells within the cell suspension can be avoided. The present inventors have developed an anti-aggregate device, as described herein. The prototype device of the present invention, as described in Example 3, has been demonstrated to be particularly effective at mixing cells and thus having an anti-aggregate effect, as shown, among others, in Example 4.

In a first aspect, the present invention relates to an anti-aggregate device, the device configured to hold an infusion bag with a liquid and deliver the liquid through the tubing, the device comprising (a) means for mixing the liquid held in the infusion bag, and (b) casing enclosing (a) and the infusion bag; wherein (a) is configured to be contacted with the infusion bag at a plurality of contact interfaces, and wherein (a) is configured to mix the liquid held in the infusion bag (a) by pressing onto the infusion bag and displacing the liquid at each contact interface, while (a) is undergoing motion such that each contact interface between (a) and the infusion bag is moving along a closed trajectory.

In a particular aspect, the anti-aggregate device relates to an embodiment, wherein (a) is attached to (b).

In a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the liquid is a cell suspension.

In a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the liquid contains nutrient(s) and/or other soluble components

In a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the motion of (a) causes a homogeneous distribution of cells, the nutrient(s) or the other soluble components in the infusion bag.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the motion of (a) causes flow of the cells, the nutrient(s) or the other soluble components inside the infusion bag away from tubing attached to the infusion bag.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the motion of (a) causes flow of the cells, the nutrient(s) or the other soluble components through the tubing attached to the infusion bag. In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the motion of (a) causes backflow of the cells, the nutrient(s) or the other soluble components from tubing to the infusion bag.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein (a) comprises a plurality of objects, each object comprising a blunt convex surface, configured to contact the infusion bag through said blunt convex surface at one of the plurality of contact interfaces.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein each object is configured for moving along its closed trajectory, and wherein the motion of (a) comprises moving each object along a closed trajectory.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein each object is a ball, an ellipsoid or a cylinder.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the objects are grouped in pairs, and wherein objects grouped in one pair are configured to move along the same closed trajectory, preferably in antiphase.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the objects are grouped in triads, and wherein objects grouped in one triad are configured to move along the same closed trajectory.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein (a) comprises a one or more motors, each motor configured to control motion of one or more objects along its/their closed trajectory(y/ies).

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the motion of (a) is magnetically controlled. In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the closed trajectory is an ellipse, preferably wherein the closed trajectory is a circular trajectory.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the closed trajectory can be changed during use.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein (b) further comprises a pressure pump.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the pressure pump is a ball pump or an electric pump.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein (b) is configured to maintain the plurality of contact interfaces between (a) and the infusion bag while the liquid is delivered out of the infusion bag.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein (a) is configured to compensate for decrease of volume of the infusion bag upon delivery of the liquid through the tubing.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein the relative position of (a) and the infusion bag is adapted to compensate for decrease of volume of (a) upon delivery of the liquid through the tubing.

In again a further particular aspect, the anti-aggregate device relates to an embodiment, wherein (b) further comprises means of controlling the temperature.

In a second aspect, the present invention relates to a anti-aggregate device of the present invention for use in the treatment, prevention and/or alleviation of symptoms of a disease. In a particular aspect, the anti-aggregate device for use of the present invention relates to an embodiment, wherein the treatment, prevention and/or alleviation of symptoms of a disease comprises infusion of a cell suspension.

In a further particular aspect, the anti-aggregate device for use of the present invention relates to an embodiment, wherein the disease is a cancer, preferably a haematological malignancy, preferably lymphoma.

In again a further particular aspect, the anti-aggregate device for use of the present invention relates to an embodiment, wherein the disease is selected from non-Hodgkin lymphoma, diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, high grade B-cell lymphoma, transformed follicular lymphoma and mantle cell lymphoma.

In again a further particular aspect, the anti-aggregate device for use of the present invention relates to an embodiment, wherein the cell suspension comprises CAR-T cells.

In again a further particular aspect, the anti-aggregate device for use of the present invention relates to an embodiment, wherein the cell suspension comprises stem cells.

In a further aspect, the present invention relates to an anti-aggregate device for use in nutrition.

Definitions

Within the meaning of the present application the following definitions apply:

“Cell suspension” is herein understood as heterogeneous mixture comprising a liquid medium, which may also be referred to as a medium, and cells suspended therein, floating in the solution. As preferably understood herein, in the absence of mixing and/or agitation the cells in the cell suspension will eventually settle and will no longer float in the solution. As understood herein, the medium is typically an aqueous solution capable of keeping the cells alive for a prolonged period of time. Suitable media are known to the skilled person.

“Aggregate” as understood herein typically refers to a non-naturally occurring cellular aggregate, that is a formed cluster of cells in the suspension that no longer free float in the suspension, but become attached to each other forming multicellular particles. Preferably, the term aggregate does not comprise aggregates that occur in a biological system. Further preferably, the term aggregate encompasses macroscopic aggregates and/or aggregates capable of interfering with function of the cells. As understood herein, the cellular aggregate as defined herein comprises a plurality of cells attached together that they float together in a suspension. However, the term aggregate may also refer to a situation wherein components of the liquid are no longer homogeneous, for example when nutrient(s) or other soluble components form particles, that preferably may block the tubing of the infusion bag.

A “plurality” preferably as used herein is meant to understand an integer value that is more than one. Therefore, the term “plurality” is preferably to be construed herein as referring to “at least two”.

A “trajectory” as understood herein is preferably defined as a path followed by a moving object. The trajectory as defined herein may be closed, also referred to as “a closed trajectory”, that is referring to a movement of an objected that returns to a starting point, and preferably continues thereon alongside the same trajectory.

A term “homogeneous”, in particular with respect to “homogeneous distribution of the cells in the cell suspension” refers to a situation where the suspension is substantially free of aggregates.

Brief description of figures

Fig. 1: The anti-aggregate device of the present invention (100), holding an infusion bag (101 ) holding a liquid (102), the bag is connected through tubing to a patient. The device comprises the means for mixing the liquid held in the infusion bag (103), and a casing (104) encompassing the means for mixing the liquid held in the infusion bag, and the infusion bag.

Fig. 2: The anti-aggregate device of the present invention, further comprising a pressure pump, i.e. a manometric device, within the casing, the pressure pump comprising the inflatable pouch (201 ) and hand-operated manometric device (202).

Fig. 3: The anti-aggregate device of the present invention - prototype. 1 . Mounting bracket L 90x90x5 3.3206 (EN-AW 6060) 2. Proto main box 6 3.3206 (EN- AW 6060), 3. PMMA carrier ball support, 4. Main shaft 3.3206 (EN-AW 6060), 5. 42SHDC3025-24B stepper motor, 6. Carrier ball 7. Bearing 6201- 2Z, 8. Actuator JF1039 12V - 10mm - 25N 1 , 9. Guidance IGUS, 9.1 Rail TS04-07 3.3206 (EN-AW 6060), 9.2 Shoe TW04-07 Nylon 101 , 10. Stop ring DIN 471 - 12 x 1 , 11. DIN 913 - M4 x 5 flat end hexagon socket grub screw, 12. Snap ring DIN 472 - 32 x 1.2, 13. Hexagonal socket head screw DIN 7991 - M3 x 12, 14. Hexagon socket head cap screw DIN 912 - M3 x 16, 15. Spring pin DIN EN 13337 - 2.5 x 22, 16. Hexagon socket head cap screw DIN 912 - M2 x 6, 17. Hexagon socket head cap screw DIN 912 - M3 x 8.

Fig. 4: Trajectories tested in the present study. Mixing head rotational speed is approximately 75 rpm for trajectories A, B and C.

Fig. 5: Experimental setup and blood count results for a single experiment of 15 minutes with an 8-shape sweep at a speed of 1500 mm/min (pool and split a).

Fig. 6: Experimental setup and blood count results for a representative measurement from a triplicate of 15 minutes with an 8-shape sweep (trajectory A, see Figure 4) at a speed of 3000 mm/min (pool & split b). Fig. 7: Experimental setup and blood count results for a single experiment of 5 minutes with an 8-shape sweep (trajectory B, see Figure 4) at a speed of 3000 mm/min (pool & split c) (part 2) and a single of 5 minutes with a up/down-shape sweep (trajectory C, see Figure 4) at a speed of 3000 mm/min (pool & split c) (part 3).

Fig. 8: Exemplary progress of mixing as shown in pictures taken every second. Pictures were taken from a representative test using trajectory C.

Detailed description of the invention

The embodiments of the present invention will be described in the following. It is to be understood that all possible combinations of the following definitions and/or features are also envisaged.

In a first aspect, the present invention relates to an anti-aggregate device. The antiaggregate device according to the present invention is configured to hold an infusion bag with a liquid and deliver the liquid through the tubing. The device comprises (a) means for mixing the liquid held in the infusion bag, and (b) casing enclosing (a) and the infusion bag. As understood within the scope of the present invention, the means for mixing the liquid held in the infusion bag (a) is/are contacted with the infusion bag at a plurality of contact interfaces. Further, the means for mixing the liquid held in the infusion bag (a) is/are configured to mix the liquid in the infusion bag by pressing onto the infusion bag and displacing the liquid at each contact interface. Further, the means for mixing the liquid held in the infusion bag (a) is/are undergoing motion such that each contact interface between the means for mixing the liquid held in the infusion bag (a) and the infusion bag is moving along a closed trajectory.

The anti-aggregate device according to the present invention is configured to hold an infusion bag with the liquid. Preferably, an infusion bag is placed in the device of the present invention so that it contacts the means for mixing the liquid held in the infusion bag (a). The means for mixing may form a structure that holds the infusion bag in place, and for example does not allow it to fall through the device, i.e. through the said means for mixing.

Optionally, in certain embodiments the anti-aggregate device of the present invention may further comprise a holder for an infusion bag. The holder may take a form of a structure comprising an attachment point, which for example may be a hook (which however is not to be construed as limiting), and an infusion bag may be attached to the holder. For example, the infusion bag may be hanging from the said attachment point, for example from the hook. Alternatively, the holder may take a form of a support structure, for example a basket, and an infusion bag may be placed therein. As noted herein, the holder is an optional feature and is not particularly limited - any holder that a skilled person can envisage as useful in the present device is considered to be encompassed within the scope of the present invention.

It is noted that the holder is to be configured so that the means for mixing the liquid held in the infusion bag (a) is/are contacted with the infusion bag, which is placed in the holder, at a plurality of contact interfaces. In certain embodiments, the holder may take a form of a bag, wherein an infusion bag is to be placed. Herein, it is to be understood that the means for mixing the liquid held in the infusion bag (a) is/are contacted with the holder, which can also be referred to as a holder bag, at a plurality of contact interfaces. As the infusion bag is to be placed in the holder bag, it is to be understood that within the scope of the present invention, the means for mixing the liquid held in the infusion bag (a) is/are contacted with the infusion bag through the holder bag at said plurality of contact interfaces.

In certain embodiments, the casing (b) may play a role of the holder. In certain embodiments, the casing (b) may be the holder. Suitable forms of the casing (b) are discussed herein.

The device of the present invention is preferably suitable for holding of any type of an infusion bag, known in the everyday practice to a skilled clinician. In other words, any type of infusion bag used in the clinical practice is preferably suitable for use with the device of the present invention. Preferably, the dimension of the means for mixing the liquid held in the infusion bag and/or dimensions of the casing (b), and/or optionally dimension of the holder may be adjusted to different types and sizes of the infusion bag. Alternatively, the device of the present invention may be so manufactured, to be configured for holding certain suitable types of an infusion bag. It is noted that skilled person will know how to adapt a design, dimensions and/or configuration, among others, of the anti-aggregate device of the present invention, to a particular type of the infusion bag. As understood herein, the infusion bag is an infusion bag with a tubing. It is to be construed as an infusion bag with integrated tubing, or an infusion bag to which tubing can be attached.

According to the present invention, the anti-aggregate device is configured to hold an infusion bag with a liquid and deliver the liquid through the tubing. As understood herein, the liquid is to be delivered from the infusion bag according to the ways and means known to the skilled person. Typically, gravity flow of the liquid out of the infusion bag through the tubing occurs.

According to the present invention, the anti-aggregate device of the present invention comprises means for mixing the liquid held in the infusion bag. The said means for mixing the liquid held in the infusion bag are not to be particularly limited from the structural point of view. Within the scope of the present invention it is required that the means for mixing the cell suspension held in the infusion bag (a) are/is configured to be contacted with the infusion bag at a plurality of contact interfaces. In other words, when the anti-aggregate device of the present invention holds the infusion bag, the means for mixing the liquid held in the infusion bag (a) are contacted with the infusion bag at a plurality of contact interfaces. The number of contact interfaces is herein not particularly limited, and may depend on the structure of the means for mixing the liquid held in the infusion bag (a). As understood herein, the contact between the means for mixing the liquid and the infusion bag holding the liquid is required to involve exerting certain force by the means for mixing the liquid onto the infusion bag, which is sufficient to affect the shape of the said infusion bag, e.g. by creating a temporary dent therein. In other words, the means for mixing the liquid held in the infusion bag (a) are configured to press onto the infusion bag and displace the liquid at a plurality of contact interfaces. Displacing the liquid, as preferably understood herein, is defined as reshaping the infusion bag upon pressing thereonto. It is herein noted that the means for mixing the liquid held in the infusion bag (a) and the infusion bag itself are not required, according to the present invention, to be in direct contact, as an additional layer of material may be placed in between to mediate the contact. It is further noted that the contact between the means for mixing the liquid held in the infusion bag and the infusion bag holding the liquid may also be referred to as pressing onto the infusion bag by the means for mixing the liquid held in the infusion bag.

Within the scope of the present invention, it is required that the means for mixing the liquid held in the infusion bag (a) are further configured to mix the liquid held in the infusion bag by pressing onto the infusion bag and displacing the liquid at a plurality of contact interfaces, preferably at each contact interface. As understood herein, within the scope of the invention the mans for mixing the liquid held in the infusion bag (a) are undergoing motion such that each contact interface between (a) and the infusion bag is moving along a closed trajectory. As the contact interfaces are moving, the shape of the infusion bag is changing, which leads to variable displacement of the liquid held in the infusion bag. As further understood herein, variable and changing with time displacement of the liquid held in the infusion bag leads to movement of the liquid within the infusion bag, which may also be referred to as flow of the liquid inside the infusion bag.

The term “motion”, as understood within the scope of the present invention is meant to comprise the motion trajectory for each contact interface between the means for mixing the liquid held in the infusion bag and the infusion bag, and the velocity(y/ies) for each contact interface (respectively, a point on the contact interface representative of the said contact interface). Preferably, the closed trajectory is an ellipse, more preferably the closed trajectory is a circular trajectory. In certain embodiments, the closed trajectory as referred to herein can be changed, for example can be changed during the infusion, as required.

Preferably, the liquid as discussed herein, is a cell suspension. Therefore, the means for mixing the liquid held in the infusion bag may also be referred to as means for mixing the cell suspension held in the infusion bag. However, in certain embodiments of the present invention, the liquid beyond the cell suspension may be encompassed by the present invention. In particular, in certain embodiments of the present invention, the liquid as defined herein contains nutrient(s) and/or other soluble components, which preferably are intended for infusing the patient.

Within the scope of the present invention, the motion of the means for mixing the liquid held in the infusion bag (a) can be so configured to cause the flow of the cells, nutrient(s) or the other soluble components inside the infusion bag. As understood to the skilled person, any aggregates that may form in the liquid (for example cell suspension, or the liquid containing nutrient(s) and/or other soluble components) would have negative impact on the infusion or the flow of the cells, nutrient(s) or other soluble components through the tubing, if they were to move into the tubing and obstruct it. Therefore, any flow of the cells, nutrient(s) or other soluble components inside the infusion bag can be defined with respect to the tubing that, as understood herein, is to be attached to the infusion bag.

Accordingly, the motion of the means for mixing the liquid held in the infusion bag (a) can be so configured to cause the flow of the cells, nutrient(s) and/or other soluble components inside the infusion bag, away from tubing. As understood herein, as the flow through the tubing is preferably, at least in part, gravity controlled, it would mean that the means for mixing the liquid held in the infusion bag (a) would have to be configured to cause flow of the cells, nutrient(s) or other soluble components inside the bag, at least in the proximity of the tubing, in the direction against gravity. In certain embodiments, the means for mixing the liquid held in the infusion bag (a) can be so configured to aid the flow of the cells, nutrient(s) or other soluble components through the tubing. In other words, the motion of the means for mixing the liquid held in the infusion bag (a) can be so configured to cause and/or to aid, preferably to cause the flow of the cells, nutrient(s) or other soluble components through the tubing. In this embodiment, in particular wherein the liquid is a cell suspension, the motion of the means mixing the liquid held in the infusion bag (a) not only prevents aggregation of the cells in the cell suspension, but also aids the delivery of the cell suspension through the tubing.

Further within the scope of the invention, the motion of the means for mixing the liquid held in the infusion bag (a) can be so configured to cause backflow of the cells, nutrient(s) or other soluble components from tubing to the infusion bag (a). As understood herein, as the flow through the tubing is preferably, at least in part, gravity controlled, it would mean that the means for mixing the liquid held in the infusion bag (a) would have to be configured to cause flow of the cells, nutrient(s) or the other soluble elements in the direction against gravity, back to the inside of the infusion bag. Such a flow would aid removing aggregates from the tubing, should any be formed already and negatively impact the delivery of the liquid through the tubing.

Further within the scope of the invention, the motion of the means for mixing the liquid held in the infusion bag (a) can be so configured to cause a homogeneous distribution of cells, nutrient(s) or other soluble components in the infusion bag. The homogeneous distribution of cells, nutrient(s) or other soluble components is understood herein as wherein substantially no cells, nutrient(s) or other soluble components are present as aggregates, as defined herein. As understood herein, in the embodiment wherein the liquid is a cell suspension, the homogeneous distribution of cells in the infusion bag can be effected by mixing the cell suspension, for example by causing the flow of the cell suspension inside the infusion bag. In certain embodiments, different contact interfaces between the means for mixing the cell suspension held in the infusion bag (a) and the infusion bag may be moving in different direction(s), such that turbulent flow of the cell suspension is caused. Turbulent flow may prevent formation of aggregates in the cell suspension, and/or disrupt the aggregates that have already formed. As understood herein, turbulent flow preferably refers to a situation wherein the fluid motion cannot be described as the motion of layers of fluid without disruption between these layers.

Within the scope of the present invention, preferably the means for mixing the liquid held in the infusion bag (a) comprise(s) a plurality of objects. It is preferably understood herein that each object of the plurality of objects is configured to be in contact with the infusion bag, that each object of the plurality of object forming an interface of the plurality of interfaces between the means for mixing the liquid held in the infusion bag (a) and the infusion bag.

The object, as referred to herein, is not particularly limited and any object suitable for the use as means of mixing the liquid held in the infusion bag, as apparent to the skilled person, can be used within the scope of the present invention. Preferably, each object of the plurality of objects of the means for mixing the liquid held in the infusion bag comprises a blunt convex surface, configured to contact the infusion bag through said blunt convex surface at one of the plurality of contact interfaces. In certain embodiments, shapes of each object of the plurality of objects may also be different, and may comprise no blunt convex surface. It is however noted that the objects comprising sharp edge and/or sharp apex, in particular configured so that the object contacts the infusion bag through said sharp edge and/or sharp apex, are not preferred in within the scope of the present invention, as their use could damage the infusion bag.

Preferably, within the scope of the present invention each object of the plurality of objects of the means for mixing the liquid held in the infusion bag can be independently selected from an object of a shape of a ball, an ellipsoid and a cylinder. In other words, each object of the plurality of objects of the means for mixing the cell suspension held in the infusion bag can comprise of or consist of a ball, an ellipsoid and a cylinder. It is understood herein each object of the plurality of objects of the means for mixing the cell suspension held in the infusion bag (a) is configured to be contacted with the infusion bag at a contact interface between said object and the infusion bag, wherein the contact interfaces described together as a plurality of contact interfaces between the means for mixing the liquid held in the infusion bag and the infusion bag. It is noted that within the scope of the present invention it is not required that each object of the plurality of objects of the means for mixing the liquid held in the infusion bag (a) have the same shape.

Preferably within the scope of the present invention each object of the plurality of objects of the means for mixing the liquid held in the infusion bag is configured for moving along a closed trajectory, in particular along its closed trajectory, wherein trajectories of different objects can be different, or can overlap. It is noted herein that as understood herein, the motion of (a) comprises moving each object along a closed trajectory.

In certain embodiments, the objects of the plurality of objects of the means for mixing the liquid held in the infusion bag (a) are grouped in pairs, which are herein referred to as pairs of objects. Accordingly, said pair of objects is configured to contact the infusion bag at a contact interface, which may be a composite contact interface comprising the contact interfaces of both objects of the pair of objects.

As understood herein, each pair of objects is configured to mix the liquid held in the infusion bag (e.g. a cell suspension) by pressing onto the infusion bag and displacing the cell suspension at each contact interface. Further, each pair of objects is undergoing motion such that each contact interface between the means for mixing the liquid held in the infusion bag (a) and the infusion bag is moving along a closed trajectory. As understood herein, the objects of each pair of objects are undergoing the motion, as described herein, in an interrelated matter. The motion of each of the object within the object pair may be in phase, or in antiphase. Preferably both objects of the pair of objects during the motion remain in the same relative position of each other on the trajectory, as expressed in a coordinate alongside the trajectory. Preferably, the pair of objects comprises two balls, or consists of two balls. Preferably, the balls are of different size. In a preferred embodiment, the trajectory is circular and the balls remain on the opposite sides of the circle with respect of each other. Further preferably, the pairs of balls as defined herein are configured to mix the liquid held in the infusion bag (e.g. a cell suspension) by pressing onto the infusion bag and displacing the liquid (e.g. the cell suspension) at each contact interface, while each ball of the pair is undergoing a circular motion, being placed on the opposite sides of the circle with respect to each other.

As understood herein, in certain embodiments of the present invention the motion of all pairs of balls considered together causes a homogeneous distribution of cells, nutrients or the other soluble components in the infusion bag. In certain embodiments of the present invention the motion of all pairs of balls considered together causes flow of the liquid (e.g. the cell suspension) inside the infusion bag away from tubing. In certain embodiments of the present invention the motion of all pairs of balls considered together causes flow of the liquid (e.g. cell suspension) through the tubing. In certain embodiments of the present invention the motion of all pairs of balls considered together causes backflow of the liquid (e.g. cell suspension) from tubing to the infusion bag.

In certain preferred embodiments of the present invention the objects of the plurality of objects of the means for mixing the liquid held in the infusion bag (a) are grouped in the groups of three, which are herein referred to as triads of objects. Accordingly, said triad of objects is configured to contact the infusion bag at a contact interface, which may be a composite contact interface comprising the contact interfaces of two or three objects of the triad of objects.

As understood herein, each triad of objects is configured to mix the liquid held in the infusion bag (e.g. a cell suspension) by pressing onto the infusion bag and displacing the cell suspension at each contact interface. Further, each triad of objects is undergoing motion such that each contact interface between the means for mixing the liquid held in the infusion bag (a) and the infusion bag is moving along a closed trajectory. As understood herein, the objects of each triad of objects are undergoing the motion, as described herein, in an interrelated matter. The motion of each of the object within the object triad may be in phase, or in antiphase. Preferably all the objects of the triad of objects during the motion remain in the same relative position of each other on the trajectory, as expressed in a coordinate alongside the trajectory.

Preferably, the triad of objects comprises three balls, or consists of three balls. Preferably, the balls are of the same size. In a preferred embodiment, the trajectory is circular and the balls remain distributed evenly across said circular trajectory with respect of each other. Further preferably, the triads of balls as defined herein are configured to mix the liquid held in the infusion bag (e.g. a cell suspension) by pressing onto the infusion bag and displacing the liquid (e.g. the cell suspension) at each contact interface, while each ball of the triad is undergoing a circular motion, being distributed evenly across said circular trajectory with respect of each other.

As understood herein, in certain embodiments of the present invention the motion of all triads of balls considered together causes a homogeneous distribution of cells, nutrients or the other soluble components in the infusion bag. In certain embodiments of the present invention the motion of all triads of balls considered together causes flow of the liquid (e.g. the cell suspension) inside the infusion bag away from tubing. In certain embodiments of the present invention the motion of all triads of balls considered together causes flow of the liquid (e.g. cell suspension) through the tubing. In certain embodiments of the present invention the motion of all triads of balls considered together causes backflow of the liquid (e.g. cell suspension) from tubing to the infusion bag.

Further encompassed by the invention are embodiments wherein the objects of the plurality of objects of the means for mixing the liquid held in the infusion bag (a) are grouped in the groups of four, five, six, seven, or more objects.

As understood within the scope of the present invention, the means for mixing the liquid held in the infusion bag (a) may in certain embodiments also comprise one or more motor(s). As understood herein, the one or more motors is configured to control the motion of the means for mixing the liquid held in the infusion bag. Accordingly, the one or more motors is configured to control the mixing of the contents of the infusion bag, in particular to control the mixing of the liquid held in the infusion bag. In certain preferred embodiments, each motor is configured to control motion of one or more object(s) along its/their closed trajectory(yZies). In an embodiment, wherein the objects of the plurality of objects are grouped in pairs, preferably each motor controls the motion of one pair of objects. In an embodiment, wherein the objects of the plurality of objects are grouped in triads, preferably each motor controls the motion of one triad of objects.

The motor as referred to herein is not particularly limited and any means of propelling the motion of the means for mixing the liquid held in the infusion bag (a) can be used within the scope of the present invention. In certain embodiments, the motor is a battery-powered electric motor. In certain embodiments, the motion of the means for mixing the liquid held in the infusion bag (a), preferably the motion of the plurality of objects (or the pairs of objects or the triads of objects), is magnetically controlled.

According to the present invention, the anti-aggregate device of the present invention further comprises the (b) casing enclosing (a). As understood herein, the casing may be further enclosing the infusion bag that is held in the anti-aggregate device of the present invention. Preferably, the means for mixing the liquid held within the infusion bag (a) is/are attached to the casing (b).

In certain embodiments, the casing (b) may be a rigid object with the means for mixing the liquid held within the infusion bag (a) rigidly attached thereto. In certain embodiments, the casing (b) may be so configured that its shape may be adjusted, and configuration of the means for mixing the liquid held within the infusion bag (a) may be adjusted accordingly, for example to accommodate the infusion bag of particular size. In certain embodiments, the casing (b) may comprise elastic material. For example, the casing may be made of cloth. In certain embodiments, the casing (b) further comprises a pressure pump. As understood herein, the pump may comprise an inflatable pouch and means for inflating the pouch. The inflatable pouch, or the pouch, can also be referred to herein as an inflatable bag. The pouch is to be placed within the casing (b) and to be configured to apply pressure onto the infusion bag. The pressure can be applied onto the infusion bag directly, or indirectly through applying pressure onto the means for mixing the liquid held in the infusion bag. In certain embodiments, the inflatable pouch may be integrated with the casing (b), or the casing (b) may be an inflatable pouch. According to the present invention, the means for inflating the pouch, within the pressure pump as understood herein, may include a hand-operated pump (which may also be referred to as manometric device) or an electric pump. Any means known to the skilled person may be used herein.

The casing (b) as defined herein, in particular the casing further comprising a pressure pump as defined herein is configured to maintain the plurality of contact interfaces between the means for mixing the liquid held within the infusion bag (a) and the infusion bag, in particular while the liquid (e.g. the cell suspension) is delivered out of the infusion bag (a) through the tubing. In certain embodiments of the present invention, the casing (b) as defined herein, in particular the casing further comprising a pressure pump as defined herein is configured to for decrease of volume of the infusion bag upon delivery of the cell suspension through the tubing, wherein the relative position of (a) and the infusion bag is adapted to compensate for decrease of volume of the infusion bag upon delivery of the liquid (e.g. the cell suspension) through the tubing. The pressure pump within the scope of the present invention may also, by applying pressure onto the infusion bag, aid delivery of the liquid (e.g. the cell suspension) out of the infusion bag.

Preferably, as encompassed by the present invention the dimension of the means for means for mixing the liquid held in the infusion bag (a) and/or dimensions of the casing (b), and/or optionally dimension of the holder may be adjusted to different types and sizes of the infusion bag. In certain embodiments, the dimension of the means for means for mixing the liquid held in the infusion bag (a) and/or dimensions of the casing (b), and/or optionally dimension of the holder may be adjustable to different types and sizes of the infusion bag.

In certain embodiments of the present invention, the casing (b) optionally further comprises the means of controlling the temperature inside the anti-aggregate device. As known to the skilled person, the contents of the infusion bag held in the device, depending on the cell suspension held in the infusion bag, may have to be temperated to maintain properties and/or stability of the cells within the cell suspension.

In another aspect, the present invention relates to use of the anti-aggregate device of the present invention in therapy. In particular, the device of the present invention is useful in infusion. As discussed herein, the infusion bag that the device is configured to hold the liquid, preferably the cell suspension. Any cell suspension, as known to the skilled person, can be used with the device of the present invention. The device of the present invention is particularly suitable for the use with cell suspension, which is prone to aggregate. In particular the present invention relates to an anti-aggregate device as described herein, wherein the cell suspension comprises CAR-T cells, wherein the cell suspension comprises tumour-infiltrating lymphocytes (TILs) or wherein the cell suspension comprises the stem cells.

In a further aspect, the present invention relates to an anti-aggregate device of the present invention for use in the treatment, prevention and/or alleviation of symptoms of a disease. Preferably, the device of the present invention is to be used for infusion of the CAR-T cells. Thus, in another aspect, the present invention relates to an antiaggregate device for use in the treatment, prevention and/or alleviation of symptoms of a disease, wherein the disease is a cancer, preferably the cancer that can be treated with CAR-T cells. Preferably, the cancer is a haematological malignancy, preferably lymphoma. In certain embodiment of the present invention, the cancer is selected from non-Hodgkin lymphoma, diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, high grade B-cell lymphoma, transformed follicular lymphoma and mantle cell lymphoma. As understood within the scope of the present disclosure, the present invention encompasses any further use of the anti-aggregate device of the present invention. In certain embodiments, the present invention relates to an anti-aggregate device of the present invention for use in parenteral nutrition.

The invention is illustrated by the following examples which, however, are not meant to be construed as limiting.

Examples

Example 1 .

The anti-aggregate device as shown in Figure 1. The device (100) comprises a slipcover (also referred to as casing 104) encompassing means for mixing a liquid (herein a cell suspension) held in the infusion bag (103). The device is put around the infusion bag (101 ) holding the cell suspension (102). Inside the slipcover, the means for mixing the cell suspension held in the infusion bag (103) are placed, encompassing 8 rotative arms (four on each side) comprising each two balls, of different sizes, each of the balls contacting the infusion bag. The rotation is controlled by battery-powered motors. By rotating in different directions, the rotative arms displace the cell suspension inside the infusion bag causing its flow inside the bag, and mixing.

Example 2.

The anti-aggregate device as shown in Figure 2. The device further encompasses a pressure pump, herein a manometric device, within the casing, the pressure pump comprising the inflatable pouch (201 ) and hand-operated manometric device (202). In the course of delivering the cell suspension to the patient, the size of the infusion bag gets smaller. The contact between the device and the infusion bag can be maintained by inflating the inflatable pouch, so that the means of mixing the cell suspension held in the infusion bag would remain in contact with the infusion bag. Example 3.

A complete overview of the machine, which constitutes a prototype of an embodiment of the present invention, is shown on the drawings in Figure 3.

Example 4.

The effect of the anti-aggregate device as described in Example 3 is presented in the following.

For each set of experiment, 12 Buffy coat of 50 mL, obtained from the Blood Transfusion Center, Lausanne, Switzerland, were pooled together to obtain a total volume of 600 mL. This total volume was split between three transfusion bags of 200 mL.

The transfusion bags were then centrifugated at 351 g for 10 minutes to obtain a non- homogeneous/sedimented solution model in the bags.

Each experiment consisted in collecting fraction of 10 mL from the transfusion bag via a stop flow method up to a total volume of 180 mL. Each fraction was then analyzed by Sysmex and the concentration of red blood cells (RBC), white blood cells (WBC) and platelets were monitored. Three fractions (6, 10 and 14) were also analyzed to determine the mortality of the cells throughout the process. According to the performed measurements based on FACS, the percentage of dead cells was between 3.2 and 7.8%, which is applicable for use of the device of the invention in the clinical setting.

The machine used during testing moves the mixing head along X-Y axes, using predefined trajectories. The mixing head includes 3 ball casters (i.e. a triad of balls) that can rotate around their axes. A compression spring is used to apply force during the mixing sequence.

The following conditions were carried out as a control: o Homogenous control: non-centrifugated transfusion bags o Non-homogeneous control: centrifugated transfusion bags (pool & Split a - control)

The effects of the machine on the homogeneity were measured by performing the following operations: o A single experiment of 15 minutes with an 8-shape sweep (trajectory A, see Figure 4) at a speed of 1500 mm/min) (non-successful) (pool & Split a) o A duplicate of 15 minutes with an 8-shape sweep (trajectory B, see Figure 4) at a speed of 3000 mm/min (pool & split b) o A duplicate of 5 minutes with an 8-shape sweep (trajectory B, see Figure 4) at a speed of 3000 mm/min (pool & split c) o A single experiment of 5 minutes with a up/down-shape sweep (trajectory C, see Figure 4) at a speed of 3000 mm/min (pool & split c)

The results are shown in Figure 5-7. Pictures from a representative experiment are shown in Figure 8.

Further embodiments of the invention are summarized in the following numbered items.

1. An anti-aggregate device, the device configured to hold an infusion bag comprising a liquid and deliver the liquid through a tubing attached to the infusion bag, the device comprising

(a) means for mixing the liquid held in the infusion bag, and

(b) a casing enclosing (a) and the infusion bag; wherein (a) is configured to be contacted with the infusion bag at a plurality of contact interfaces, and wherein (a) is configured to mix the liquid held in the infusion bag by pressing onto the infusion bag and displacing the liquid at each contact interface, while (a) is undergoing motion such that each contact interface between (a) and the infusion bag is moving along a closed trajectory. The anti-aggregate device of item 1 , wherein (a) is attached to (b). The anti-aggregate device of item 1 or 2, wherein the liquid is a cell suspension. The anti-aggregate device of item 1 or 2, wherein the liquid contains nutrient(s) and/or other soluble components. The anti-aggregate device of item 3 or 4, wherein the motion of (a) causes a homogeneous distribution of the cells, the nutrient(s) or the other soluble components in the infusion bag. The anti-aggregate device of any one of items 1 to 5, wherein the motion of (a) causes flow of the cells, the nutrient(s) or the other soluble components inside the infusion bag away from the tubing attached to the infusion bag. The anti-aggregate device of any one of items 1 to 5, wherein the motion of (a) causes flow of the cells, the nutrient(s) or the other soluble components through the tubing attached to the infusion bag. The anti-aggregate device of any one of items 1 to 5, wherein the motion of (a) causes backflow of the cells, the nutrient(s) or the other soluble components from tubing to the infusion bag. The anti-aggregate device of any one of items 1 to 8, wherein (a) comprises a plurality of objects, each object comprising a blunt convex surface, configured to contact the infusion bag through said blunt convex surface at one of the plurality of contact interfaces. The anti-aggregate device of item 9, wherein each object is configured for moving along its closed trajectory, and wherein the motion of (a) comprises moving each object along a closed trajectory. The anti-aggregate device of item 9 or 10, wherein each object is a ball, an ellipsoid or a cylinder. The anti-aggregate device of any one of items 9 to 11 , wherein the objects are grouped in pairs, and wherein objects grouped in one pair are configured to move along the same closed trajectory, preferably in anti-phase. The anti-aggregate device of any one of items 1 to 12, wherein (a) comprises a one or more motors, each motor configured to control motion of one or more objects along its/their closed trajector(y/ies). The anti-aggregate device of any one of items 1 to 13, wherein the motion of (a) is magnetically controlled. The anti-aggregate device of any one of preceding items, wherein the closed trajectory is an ellipse, preferably wherein the closed trajectory is a circular trajectory. The anti-aggregate device of any one of preceding items, wherein the closed trajectory can be changed during use. The anti-aggregate device of any one of preceding items, wherein (b) further comprises a pressure pump. The anti-aggregate device of item 17, wherein the pressure pump is a ball pump or an electric pump. The anti-aggregate device of item 17 or 18, wherein (b) is configured to maintain the plurality of contact interfaces between the infusion bag and (a) while the liquid is delivered out of the infusion bag. The anti-aggregate device of any one of items 17 to 19, wherein (b) is configured to compensate for decrease of volume of the infusion bag upon delivery of the liquid through the tubing. The anti-aggregate device of any one of items 17 to 20, wherein the relative position of (a) and the infusion bag is adapted to compensate for decrease of volume of the infusion bag upon delivery of the liquid through the tubing. The anti-aggregate device of any one of items 17 to 21 , wherein (b) further comprises means of controlling the temperature. The anti-aggregate device of any one of items 1 to 22 for use in the treatment, prevention and/or alleviation of symptoms of a disease. The anti-aggregate device of any one of items 1 to 22 for use in nutrition. The anti-aggregate device for use of item 23, wherein the treatment, prevention and/or alleviation of symptoms of a disease comprises infusion of a cell suspension. The anti-aggregate device for use of item 23 or 25, wherein the disease is cancer, preferably a haematological malignancy, preferably lymphoma. The anti-aggregate device for use of item 26, wherein the disease is selected from non-Hodgkin lymphoma, diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, high grade B-cell lymphoma, transformed follicular lymphoma and mantle cell lymphoma The anti-aggregate device for use of item 25, wherein the cell suspension comprises CAR-T cells. 29. The anti-aggregate device for use of item 25, wherein the cell suspension comprises stem cells.

Further examples and embodiments of the present invention are disclosed in the following numbered clauses.

1. An anti-aggregate device, the device configured to hold an infusion bag comprising a liquid and deliver the liquid through a tubing attached to the infusion bag, the device comprising:

(a) means for mixing the liquid held in the infusion bag, and

(b) a casing enclosing (a) and the infusion bag; wherein (a) is configured to be contacted with the infusion bag at a plurality of contact interfaces, and wherein (a) is configured to mix the liquid held in the infusion bag by pressing onto the infusion bag and displacing the liquid at each contact interface, while (a) is undergoing motion such that each contact interface between (a) and the infusion bag is moving along a closed trajectory.

2. The anti-aggregate device of clause 1 , wherein (a) is attached to (b).

3. The anti-aggregate device of clause 1 or 2, wherein the liquid is a cell suspension or wherein the liquid contains nutrient(s) and/or other soluble components, preferably wherein the motion of (a) causes a homogeneous distribution of the cells, the nutrient(s) or the other soluble components in the infusion bag.

4. The anti-aggregate device of any one of clauses 1 to 3, wherein the motion of (a) causes flow of the cells, the nutrient(s) or the other soluble components inside the infusion bag away from the tubing attached to the infusion bag, or wherein the motion of (a) causes flow of the cells, the nutrient(s) or the other soluble components through the tubing attached to the infusion bag, or wherein the motion of (a) causes backflow of the cells, the nutrient(s) or the other soluble components from tubing to the infusion bag. The anti-aggregate device of any one of clauses 1 to 4, wherein (a) comprises a plurality of objects, each object comprising a blunt convex surface, configured to contact the infusion bag through said blunt convex surface at one of the plurality of contact interfaces, preferably wherein each object is configured for moving along its closed trajectory, and wherein the motion of (a) comprises moving each object along a closed trajectory. The anti-aggregate device of clause 5, wherein each object is a ball, an ellipsoid or a cylinder. The anti-aggregate device of clause 5 or 6, wherein the objects are grouped in pairs, and wherein objects grouped in one pair are configured to move along the same closed trajectory, preferably in anti-phase. The anti-aggregate device of any one of clauses 1 to 7, wherein (a) comprises a one or more motors, each motor configured to control motion of one or more objects along its/their closed trajector(y/ies), and/or wherein the motion of (a) is magnetically controlled, and/or wherein the closed trajectory is an ellipse, preferably wherein the closed trajectory is a circular trajectory, and/or wherein the closed trajectory can be changed during use. The anti-aggregate device of any one of preceding clauses, wherein (b) further comprises a pressure pump, and/or wherein the pressure pump is a ball pump or an electric pump, and/or wherein (b) is configured to maintain the plurality of contact interfaces between the infusion bag and (a) while the liquid is delivered out of the infusion bag, and/or wherein (b) is configured to compensate for decrease of volume of the infusion bag upon delivery of the liquid through the tubing, and/or wherein the relative position of (a) and the infusion bag is adapted to compensate for decrease of volume of the infusion bag upon delivery of the liquid through the tubing. The anti-aggregate device of clause 9, wherein (b) further comprises means of controlling the temperature. The anti-aggregate device of any one of clauses 1 to 10 for use in the treatment, prevention and/or alleviation of symptoms of a disease. The anti-aggregate device of any one of clauses 1 to 10 for use in nutrition. The anti-aggregate device for use of clause 11 , wherein the treatment, prevention and/or alleviation of symptoms of a disease comprises infusion of a cell suspension. The anti-aggregate device for use of clause 11 or 13, wherein the disease is cancer, preferably a haematological malignancy, preferably lymphoma, preferably selected from non-Hodgkin lymphoma, diffuse large B-cell lymphoma, primary mediastinal B-cell lymphoma, high grade B-cell lymphoma, transformed follicular lymphoma and mantle cell lymphoma. The anti-aggregate device for use of clause 13, wherein the cell suspension comprises CAR-T cells or wherein the cell suspension comprises stem cells.