WO2019198091A1

WO2019198091A1 - Blood based treatment

Info

Publication number: WO2019198091A1
Application number: PCT/IL2019/050418
Authority: WO
Inventors: Uri Alon; Omer Karin
Original assignee: Yeda Research And Development Co. Ltd.
Priority date: 2018-04-12
Filing date: 2019-04-12
Publication date: 2019-10-17

Abstract

The invention relates to a method for increasing blood volume in a body of a subject, comprising: attaching a container with a volume of at least 25% of the volume of the circulatory system of said subject to the circulatory system of said subject; allowing the flow of blood from said body to said container; and allowing the flow from said container back to said body; thereby increasing the blood volume of said circulatory system of said subject.

Description

BLOOD BASED TREATMENT

RELATED APPLICATION/S

This application claims the benefit of priority of U.S. Provisional Patent Application No. 62/656,413 filed on 12 April 2018, the contents of which are incorporated herein by reference in their entirety.

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to treating damage-related conditions, enhancing recovery, rejuvenation and/or for increased longevity; more particularly, but not exclusively, to treating damage-related conditions taking into account repair and/or damage processes, influencing the ratio between them and/or to increasing blood volume in a homeostatic manner.

Additional background art includes US patent application US20020052571A1, disclosing "an artificial kidney, adapted to be worn externally proximate to a patient's body, comprising, at least one blood inlet conduit, at least one micro fluidic filtering means, a means for supporting said filtering means, at least one blood outlet conduit and at least one blood solute outlet conduit".

US patent US4354933A, disclosing "an artificial kidney adapted to be implanted in the human body comprising a housing including a diffusion chamber and a dialysate bath chamber. A plurality of microtubules, preferably torqued or spiraled, are arranged substantially vertically in the diffusion chamber. Each microtubule has at least three longitudinal passages therethrough, an arterial passage, a venous passage, and at least one urinary passage, one of the urinary passages extending co-extensively between the arterial and the venous passages, and being separated by walls permeable to waste products from the bloodstream. The arterial and venous blood from the body flow counter-currently through each microtubule, so that the chemical imbalance between the arterial bloodstream and the venous bloodstream causes diffusion across the urinary passage, and the waste products are collected in, and discharged from, the urinary passage. Venous blood from the body is first transported through the dialysate bath chamber for chemical enrichment prior to its flow through the venous passages in the microtubules. The only external function required for operating the kidney is the occasional re-supply of dialysate solution into the dialysate bath". US patent US4769037A, disclosing "an artificial replacement kidney implant for dializing blood within a human body comprising at least one blood panel having an inlet for coupling to the artery of a body and an outlet for coupling to the vein of a body to receive blood to be dialized and to discharge dialized blood respectively. The blood panel includes a semi-permeable membrane through which biologically active material contained within the body of blood diffuses. An absorbent panel is disposed at the opposite side of the membrane for passing the biologically active material and mechanism is provided for communicating the biologically active material to the ureter of the body. One aspect of the invention also includes the insitu process of dializing body blood".

SUMMARY OF THE INVENTION

An aspect of the present invention is a method for increasing blood volume in a body of a subject, comprising: a. attaching a container with a volume of at least 25% of the volume of the circulatory system of said subject to the circulatory system of said subject; b. allowing the flow of blood from said body to said container; and c. allowing the flow from said container back to said body; thereby increasing the blood volume of said circulatory system of said subject.

In some embodiments, at the initial moment of attaching, said container is filled with at least one liquid other than blood.

In some embodiments, said attaching is from outside said body. In some embodiments, said attaching is from inside said body. In some embodiments, said allowing the flow comprises controlling said flow by a valve.

In some embodiments, the method further comprises providing said subject with at least one drug. In some embodiments, the drug is selected from the group consisting of: senolytic drug, hormones, vasodilatation drugs, pro-angiogenic drugs, antibiotics, steroids, anticoagulants, and any combination thereof.

In some embodiments, said method further comprises monitoring homeostatic parameters in said subject’s circulatory system.

In some embodiments, said allowing the flow comprises allowing the flow to flow in a continuous manner.

In some embodiments, said allowing the flow comprises allowing the flow to flow in an intermittent manner.

In some embodiments, said attaching is performed for a period of time of at least one week. An aspect of the present invention is a device for increasing blood volume in the body of a subject, comprising: a. at least one first tube having a first extremity and a second extremity; said first extremity interconnected in continuous flow to the circulatory system of a subject and said second extremity to at least one container; b. at least one container operatively interconnected to said second extremity of said at least one first tube; said at least one container adapted to contain at least one liquid; c. at least one second tube having a third extremity and a fourth extremity; said third extremity operatively interconnected to said container; said fourth extremity interconnected in continuous flow to said circulatory system of said subject; characterized in that the flow from said circulatory system through said container back to said circulatory system is a low flow; characterized in that said flow from said circulatory system through said container back to said circulatory system does not change the homeostatic state of said circulatory system; and characterized in that said at least one container is adapted for safely containing the blood of said subject.

In some embodiments, the device further comprises means to deliver at least one anticoagulant into said at least one first tube.

In some embodiments, the device further comprises means to deliver at least one antidote to said anticoagulant into said at least one second tube.

In some embodiments, the device further comprises means to monitor said homeostatic state of said circulatory system inside said device.

In some embodiments, the device further comprises at least one sensor.

In some embodiments, said at least one sensor is selected from the group consisting of: blood pressure, bubble sensor, arterial blood sensor, blood coagulation sensor, pH sensor, central venous sensor, oxygen sensor, cardiac output sensor, temperature sensor, and any combination thereof.

In some embodiments, the device further comprises a de-aeration chamber.

In some embodiments, the device further comprises a two-way port into said container; said two-way port interconnected to at least one third tube.

In some embodiments, the device further comprises means of oxygenation of said at least one liquid.

In some embodiments, the device further comprises means for monitoring the correct functionality of said device.

In some embodiments, the device further comprises means for controlling the temperature of said at least one liquid. In some embodiments, the device further comprises at least one power source.

In some embodiments, the device further comprises at least one controller.

In some embodiments, the device further comprises at least one graphic unit interface.

In some embodiments, the device is an external device. In some embodiments, the device an internal device. In some embodiments, the device a portable device.

An aspect of the present invention is a method of choosing a treatment for a subject, comprising:

a. evaluating a frailty score of a subject using a computer comprising circuit adapted to perform tasks according to software instructions; said evaluating comprising:

i. inserting data into said computer regarding at least one parameter selected from the group consisting of: damage levels, repair levels, current state of said subject, and any combination thereof;

ii. receiving said frailty score according to a frailty assessment model;

b. choosing a treatment according to said frailty score.

In some embodiments, when said frailty score is lower than a predetermined frailty score value, then providing pre-treatment to increase said frailty score above said predetermined frailty score value.

In some embodiments, said frailty assessment model is based on at least one model selected from the group consisting of: Linda Fried / Johns Hopkins Frailty Criteria model; Rockwood Frailty Index model; Four domains of frailty model; SHARE Frailty Index model; and any combination thereof.

An aspect of the present invention is a method of affecting a frailty status of a subject according to a frailty score assessment tool, comprising:

ii. receiving said frailty score according to a frailty assessment model; b. when said frailty score is lower than a predetermined frailty score value, then treat with a device for increasing blood volume in the body of a subject, comprising: a. at least one first tube having a first extremity and a second extremity; said first extremity interconnected in continuous flow to the circulatory system of a subject and said second extremity to at least one container; b. at least one container operatively interconnected to said second extremity of said at least one first tube; said at least one container adapted to contain at least one liquid; c. at least one second tube having a third extremity and a fourth extremity; said third extremity operatively interconnected to said container; said fourth extremity interconnected in continuous flow to said circulatory system of said subject; characterized in that the flow from said circulatory system through said container back to said circulatory system is a low flow; characterized in that said flow from said circulatory system through said container back to said circulatory system does not change the homeostatic state of said circulatory system; and characterized in that said at least one container is adapted for safely containing the blood of said subject; as described above.

Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

As will be appreciated by one skilled in the art, some embodiments of the present invention may be embodied as a system, method or computer program product. Accordingly, some embodiments of the present invention may take the form of an entirely hardware embodiment, an entirely software embodiment (including firmware, resident software, micro code, etc.) or an embodiment combining software and hardware aspects that may all generally be referred to herein as a“circuit,”“module” or“system.” Furthermore, some embodiments of the present invention may take the form of a computer program product embodied in one or more computer readable medium(s) having computer readable program code embodied thereon. Implementation of the method and/or system of some embodiments of the invention can involve performing and/or completing selected tasks manually, automatically, or a combination thereof. Moreover, according to actual instrumentation and equipment of some embodiments of the method and/or system of the invention, several selected tasks could be implemented by hardware, by software or by firmware and/or by a combination thereof, e.g., using an operating system.

For example, hardware for performing selected tasks according to some embodiments of the invention could be implemented as a chip or a circuit. As software, selected tasks according to some embodiments of the invention could be implemented as a plurality of software instructions being executed by a computer using any suitable operating system. In an exemplary embodiment of the invention, one or more tasks according to some exemplary embodiments of method and/or system as described herein are performed by a data processor, such as a computing platform for executing a plurality of instructions. Optionally, the data processor includes a volatile memory for storing instructions and/or data and/or a non-volatile storage, for example, a magnetic hard-disk and/or removable media, for storing instructions and/or data. Optionally, a network connection is provided as well. A display and/or a user input device such as a keyboard or mouse are optionally provided as well.

Any combination of one or more computer readable medium(s) may be utilized for some embodiments of the invention. The computer readable medium may be a computer readable signal medium or a computer readable storage medium. A computer readable storage medium may be, for example, but not limited to, an electronic, magnetic, optical, electromagnetic, infrared, or semiconductor system, apparatus, or device, or any suitable combination of the foregoing. More specific examples (a non-exhaustive list) of the computer readable storage medium would include the following: an electrical connection having one or more wires, a portable computer diskette, a hard disk, a random access memory (RAM), a read-only memory (ROM), an erasable programmable read-only memory (EPROM or Flash memory), an optical fiber, a portable compact disc read-only memory (CD-ROM), an optical storage device, a magnetic storage device, or any suitable combination of the foregoing. In the context of this document, a computer readable storage medium may be any tangible medium that can contain or store a program for use by or in connection with an instruction execution system, apparatus, or device.

A computer readable signal medium may include a propagated data signal with computer readable program code embodied therein, for example, in baseband or as part of a carrier wave. Such a propagated signal may take any of a variety of forms, including, but not limited to, electro-magnetic, optical, or any suitable combination thereof. A computer readable signal medium may be any computer readable medium that is not a computer readable storage medium and that can communicate, propagate, or transport a program for use by or in connection with an instruction execution system, apparatus, or device.

Program code embodied on a computer readable medium and/or data used thereby may be transmitted using any appropriate medium, including but not limited to wireless, wireline, optical fiber cable, RF, etc., or any suitable combination of the foregoing. Computer program code for carrying out operations for some embodiments of the present invention may be written in any combination of one or more programming languages, including an object-oriented programming language such as Java, Smalltalk, C++ or the like and conventional procedural programming languages, such as the "C" programming language or similar programming languages. The program code may execute entirely on the user's computer, partly on the user's computer, as a stand-alone software package, partly on the user's computer and partly on a remote computer or entirely on the remote computer or server. In the latter scenario, the remote computer may be connected to the user's computer through any type of network, including a local area network (LAN) or a wide area network (WAN), or the connection may be made to an external computer (for example, through the Internet using an Internet Service Provider).

Some embodiments of the present invention may be described below with reference to flowchart illustrations and/or block diagrams of methods, apparatus (systems) and computer program products according to embodiments of the invention. It will be understood that each block of the flowchart illustrations and/or block diagrams, and combinations of blocks in the flowchart illustrations and/or block diagrams, can be implemented by computer program instructions. These computer program instructions may be provided to a processor of a general purpose computer, special purpose computer, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, create means for implementing the functions/acts specified in the flowchart and/or block diagram block or blocks.

These computer program instructions may also be stored in a computer readable medium that can direct a computer, other programmable data processing apparatus, or other devices to function in a particular manner, such that the instructions stored in the computer readable medium produce an article of manufacture including instructions which implement the function/act specified in the flowchart and/or block diagram block or blocks.

The computer program instructions may also be loaded onto a computer, other programmable data processing apparatus, or other devices to cause a series of operational steps to be performed on the computer, other programmable apparatus or other devices to produce a computer implemented process such that the instructions which execute on the computer or other programmable apparatus provide processes for implementing the functions/acts specified in the flowchart and/or block diagram block or blocks. Some of the methods described herein are generally designed only for use by a computer and may not be feasible or practical for performing purely manually, by a human expert. A human expert, who wanted to manually perform similar tasks, might be expected to use completely different methods, e.g., making use of expert knowledge and/or the pattern recognition capabilities of the human brain, which would be vastly more efficient than manually going through the steps of the methods described herein.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

Some embodiments of the invention are herein described, by way of example only, with reference to the accompanying drawings. With specific reference now to the drawings in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention. In this regard, the description taken with the drawings makes apparent to those skilled in the art how embodiments of the invention may be practiced.

In the drawings:

Figure 1 is a schematic graph showing the relation between age, damage and repair in a subject, in accordance with some embodiments of the invention;

Figure 2 is a schematic graph showing the effects of treatments on the relation between repair factors and damage factors, in accordance with some embodiments of the invention;

Figure 3 is an illustration showing the mechanism of removal of damage factors and senescent cells by repair factors, in accordance with some embodiments of the invention;

Figure 4 is a schematic illustration showing one principle of division of the available repair resources, in accordance with some embodiments of the invention;

Figure 5 is a schematic illustration of the principle of one example of an embodiment of the present invention, in accordance with some embodiments of the invention;

Figure 6 is a schematic graph showing the expected lifespan of a subject depending on the treatment received, in accordance with some embodiments of the invention;

Figure 7 is a schematic graph showing the relation between health status and time of recovery in different conditions, in accordance with some embodiments of the invention;

Figure 8 is graphs showing the effect of different embodiments of treatment on senescent cells, in accordance with some embodiments of the invention;

Figure 9 is graphs showing the effect of different embodiments of treatment on rejuvenation state, in accordance with some embodiments of the invention; Figure 10 is graphs showing the effect of different embodiments of treatment on rejuvenation state after stop of treatment, in accordance with some embodiments of the invention;

Figures l la-c are schematic representations of different examples of some embodiment devices, in accordance with some embodiments of the invention;

Figure 12 is a schematic representation of one example of an embodiment device, in accordance with some embodiments of the invention;

Figure 13 is a schematic representation a possible connection to the circulatory system, in accordance with some embodiments of the invention;

Figure 14 is a schematic flowchart of an example of a method followed by the one embodiment of a device, in accordance with some embodiments of the invention;

Figure 15 is a schematic flowchart of some examples of monitoring performed during the activation of the device, in accordance with some embodiments of the invention;

Figure 16 is a schematic representation of an example of an embodiment device, in accordance with some embodiments of the invention;

Figure 17 is a schematic representation of an example of an embodiment device, in accordance with some embodiments of the invention;

Figure 18 is a schematic representation of an example of an embodiment device, in accordance with some embodiments of the invention; and

Figure 19 is a schematic flow chart of an example of the method followed by a physician, in accordance with some embodiments of the invention.

DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION OVERVIEW

The present invention, in some embodiments, relates to a model which helps explain the relation between age and/or the status of a subject's body with the quantity of damaging factors found in the body over time, the quantity of repair factors in the body over time and/or the effect of the relation between damage and repair over time. The model is used to make predictions about the behavior of the repair systems, and/or also help understand past behavior of the same.

A broad aspect of some embodiments of the present invention relates to generating a model of the relation between damage, repair and the current and/or potential state of a subject's body. The model uses the data related to the accumulation of damage factors over time and/or changes in the physiological condition of the subject and/or the reduction in repair capacity and interrelates between them over the time line. An aspect of some embodiments of the present invention relates to treatment methods which are based on the products from the model.

In some embodiments, products from the model are affected by modification of parameters in the model in order to help choose the method of treatment. In some embodiments, some of the parameters are modified while other are kept constant. In some embodiments, the parameters that are changed in the model are the current and/or potential state of a subject, the current and/or potential repair levels in the subject, the current and/or damage levels in the subject, the current and/or potential drugs taken by the subject.

In some embodiments, the method of treatment is used to change the actual state of the subject's body by influencing and/or maintaining at least one component in the body of the subject based on the products from the model.

In some embodiments, the method of treatment influences the damage levels and may also influence the repair levels and/or influences the repair levels and may also influence the damage levels.

In some embodiments, the methods of treatment comprise different durations of treatment (e.g. days, weeks and/or months) and different frequencies of activation (e.g.: two days“on” - one day“off’ for the duration of the treatment). In some embodiments, the methods of treatment are applied on/off for a few hours (e.g.: two hours“on” - two hours“off’ for the duration of the treatment). In some embodiments, the methods of treatment are applied from a few days to about a few months to about a few years, according to the results of the treatment over time.

In some embodiments, the method of treatment is modified according to the age of the subject and/or the state of the subject's body and/or sudden trauma.

An aspect of some embodiments of the present invention relates to modifying the volume of a blood component in a subject to change the actual state of the subject's body.

In some embodiments, the volume of at least one blood component is increased.

In some embodiments, the volume of at least one component is decreased in the subject.

In some embodiment, modifying the blood volume of at least one blood component comprises modifying while keeping a healthy homeostatic level (e.g.: within the normal parameters of the specific user) of the circulatory system.

In some embodiments, concentrations or numbers of blood cells are increased or decreased while possibly also the blood volume is increased or decreased. In some embodiments, the blood volume is increased or decreased while possibly also increasing or decreasing blood cell concentrations. In some embodiments, the volume is increased or decreased using exogenous systems (e.g., devices). In some embodiments, the volume is increased or decreased using endogenous systems (e.g., using drugs to affect body's systems).

In some embodiments, the blood component is increased or decreased using different ways, for example: using internal devices, external devices or control devices.

In some embodiments, the flow of the additional or the subtracted blood component to the subject's body is characterized by having minimal flow. In some embodiments, the flow can be controlled by gravity, by corporeal blood pressure, by compression of skeletal muscle or by an artificial pump. In some embodiments, the flow of the additional or the subtracted blood component is adapted to support long-term blood flow without clotting or obstruction.

In some embodiments, the subject can carry the device. In some embodiments, the device is used in static position (e.g.: not moved or carried around), and the subject is treated near the device.

In some embodiments, the devices are filled with blood. In some embodiments, the devices are filled with liquids other than blood. In some embodiments, the devices are filled with either saline, artificial blood, or other blood compatible liquids. In some embodiments, the liquids inside the device are in constant movement. In some embodiments, the liquids can stay static in the devices.

In some embodiments, the devices increase the blood volume while keeping the homeostatic balance in the circulatory system.

An aspect of some embodiments of the present invention relate to using the model to interrelate and estimate the actual effect of other treatment methods (e.g., drugs) in the recovery of the subject's body.

An aspect of the present invention relates to evaluating and quantifying the frailty status of a subject.

In some embodiments, the frailty status of the subject is the indication of the treatment that will be given to said subject.

An aspect of some embodiments of the present invention relates to diagnostic methods based on the products from the model.

In some embodiments, the diagnostic methods are applied for diagnosing past, present and future health states in a subject's body.

An aspect of some embodiments of the invention relates to removal of senescent cells in corporeal blood by providing the body with reactivated NK cells. In some embodiments, blood comprising exhausted NK cells is taken from a subject. In some embodiments, the blood is then incubated outside the body of the subject for a period of time. In some embodiments, the blood is incubated for a period from about 6 hours to about 24 hours. In some embodiments, the incubation of the blood outside the body allows the reactivation of exhausted NK cells. In some embodiments, after the incubation period, the blood is returned to the subject. In some embodiments, return of incubated blood is followed by a removal of new blood from the subject. In some embodiments, removal of new blood is followed by return of incubated blood. In some embodiments, small amounts of blood are taken from the subject during the day and then small amounts of incubated blood are returned after the incubation period.

Exemplary model - damage repair

It has been disclosed, for example by H. Vielle (“Aging 101: Biological causes of aging”, January 2017), that there is a tight relation between damage factors, repair factors and efficiency of repair. This relation can be described, for example, as disclosed in the schematic graph 100 in Fig. 1, presenting the relation between the accumulations of damage factors 102 over time, the quantity of repair factors 104 over time and the efficiency of repair 106 over time. Over years, the body accumulates damage factors 102. These factors originate from different sources, like trauma, natural deterioration, exposure to elements, accumulation of senescent cells and others. At the same time, repair factors 104 are naturally manufactured by the body. Over the years, the mechanisms of manufacture of repair factors naturally decline. The relation between the accumulation of damage factors and the manufacture (quantity) of repair factors is graphically showed as the efficiency of the repair 106. It has been proved that the efficiency of repair 106 declines over the years. It is suspected that one of the reasons of this decline is also due to the accumulation of damage factors and the inability of the repair factors to overcome and eliminate said damage factors and the cells from where they are originated, and the saturation of repair capacity by the increasing levels of damage production with age.

In some embodiments, the relation between repair factors and damage factors over time can be illustrated in a quantitative way, for example, as shown in the schematic illustration 200 in Fig. 2. The top graph 202 shows the accumulation of damage factors and the manufacture of repair factors over the years in a regular person (Untreated). The bars show how much damage factors accumulate in the body (showed as black bars) in a regular subject as he ages, while the quantity of repair factors remain roughly the same (showed as white bars). As disclosed above, it is known that over the years, damaging factors accumulate in the body and repair factors work to eliminate them. While young subjects manage to eliminate more efficiently these damage factors, old subjects do not. Therefore, the increase in the number of damage factors, together with less efficient repair systems, is thought to be one of the mechanisms of ageing.

In some embodiments, integration of experimental results are taken and used for developing a model for estimating damage accumulation and repair during aging. In some exemplary embodiments, data for the model is taken from measurements of senescent cell accumulation in mice, primates and human, including whole-body luminescence measurements of senescent cell dynamics in adult mice, and measurements of senescent cell accumulation in individual tissues in mice, primates and human. In some embodiments, other data for the model is recovered from heterochronic parabiosis in mice. In some embodiments, further data for the model comes from experiments on removal of senescent cells in mice by pharmaco-genetic and pharmacological means, both systemically and locally. In addition, in some embodiments, calibration of the model is performed using statistics on morbidity, frailty and mortality in mice and humans. In some embodiments, the model is used to evaluate and estimate the state of damage and repair in a subject depending on the physical status, the age and possible treatments. Using Fig. 2 as an example, one scope of the present invention is to enable a state were the damage factors (black bars) showed in 202 are smaller than the repair factors (white bars).

Exemplary model - senescent cells

In some embodiments, another marker of ageing is the accumulation of senescent cells. Recent reports show that killing senescent cells in mice increases lifespan by a third and reduces morbidity (including cancer prevalence, heart and kidney dysfunction). It is thought that the reason for this is because while senescent cells do not divide anymore, they continue to secrete factors that are damaging, such as factors causing inflammation, reducing regeneration and causing tissue lesions. The same repair factors are busy eliminating senescent cells over the years. This process is explained, for example, in the schematic representation 300 in Fig. 3. In general, senescent cells 302 begin to appear in the body and repair mechanisms 304 begin eliminating them. The senescent cells secrete damage factors 306 while they keep accumulating. Usually, in young populations, the repair factors 304 succeed in eliminating the damage factors 306 while eliminating the senescent cells 302 as well. The problem begins where repair factors cannot overcome both the senescent cells and the damage factors. This creates two individual problems: on one side, the accumulation of senescent cells and the defective regenerative processes; and on the other side, the accumulation of damage factors.

The problem of the accumulation of senescent cells, the defective regenerative processes and the accumulation of damage factors, can be explained, for example, as described in the schematic illustration 400 in Fig. 4. Generally, each person has a determined quantity of repair resources at his/her disposal 402. In some cases, a high accumulation of senescent cells and the secretion of damage factors from them 404, cause a high quantity of repair resources 406 to be invested in the elimination of the senescent cells and the damage factors. If by any chance, the same subject gets sick, by influenza 408 for example, less repair resources 410 are available to deal with this sickness. And, if another incident happens, for example, if they fall and fracture bones 411, then even less repair resources are available 412, causing a severe delay in the recovery.

Exemplary model - senolytics

The discovery of the relation between senescent cells, repair and aging encouraged the search for drugs that can kill senescent cells in humans (senolytics). While this search is still in progress, it has shown some problems of side effects in subjects. Returning to Fig. 2, in the second graph 204, marked as "Senolytics", shows how the use of these drugs can eliminate the senescent cells in the body to some degree. Further information on senolytics methods can be found for example at WO2015116740A1, incorporated herein by reference in its entirety.

Exemplary principle of treatment

In some embodiments, one of the principals of the model is to enable the evaluation of the ratio between the damage and the repair. In some embodiment of the present invention, the scope is to treat a subject in a way that the subject's damage level will be reduced by increasing their repair to damage ratio. Referring now to Fig. 5, a schematic illustration 500 of one example of an embodiment of treatment of the present invention is shown. In some embodiments, the concept is to increase the repair by increasing the subject's total blood volume, while keeping blood component concentrations and blood pressure at their homeostatic level, or at least at a healthy level compared to the healthy level of the specific user. As shown in Fig. 5, the circulatory system 502 is directly connected 504 to a container 506 having blood (or other adequate liquid), and then from the container 506, directly connected 508 back to the subject’s 510 circulatory system 502. Different examples of embodiments of devices for performing this will be further described below.

In some embodiments, the concept behind this action is that when the blood volume of the subject is increased, by example, by a factor of 1.5 (e.g.: from 4 liters to 6 liters), the natural repair mechanisms are capable to sense this increment in volume, or rather, the dilution of the repair factors relative to the total volume of blood, therefore causing an increase in the manufacture of repair blood-borne factors (including NK cells, monocytes, T cells that remove senescent cells). In some embodiments, the increment in repair factors in the blood enables the body to eliminate senescent cells and damage factors, which are also diluted due to the increase in the blood volume. In some embodiments, the more repair factors, the more elimination of damage factors. In some embodiments, the decrease in damage factors allows more repair factors to be free to deal with other repair processes (other than elimination of damage factors). Returning to Fig. 2, the third graph 206, shows how, in some embodiments, the increase in the volume of the blood will cause an increase in the repair factors (white bars) over time, which will cause a reduction in the damage factors (black bars).

In some embodiments of the present invention, further to increasing the subject's total blood volume, while keeping blood component concentrations and blood pressure at their homeostatic level, a drug (e.g. senolytic drug, anti-inflammatory drugs, antibiotics) is provided to the subject. Further information on senolytics methods can be found for example at W02015116740A1, incorporated herein by reference in its entirety. As shown in Fig. 2, the fourth graph 208, schematically shows the synergistic effect of increasing the blood volume and providing drug to subject. The effect is seen as the further reduction of damaging factors (e.g.: senescent cells) while increasing the repair factors.

Exemplary relation between lifespan and treatment

Referring now to Fig. 6, a graph 600 showing the expected lifespan of a subject depending on the treatment received. In some embodiments, according to the model data, untreated subjects have an average lifespan expectancy of 80 years 602. In some embodiments, according to the model data, subjects subjected to senolytic treatment will have an average lifespan expectancy of about 109 years 604. In some embodiments, according to the model data, subjects treated with the device of the present invention, having a 25% increased blood volume will have an average lifespan expectancy of about 102 years 606. In some embodiments, according to the model data, subjects treated with the device and with senolytics will have an average lifespan expectancy of about 137 years 608, showing the synergistic effect of the combined treatment.

In some embodiments, the model allows to calculate the senescent cell content of a human body given a time-dose course of senolytic drugs that kill a specified fraction of the senescent cells.

Exemplary treatment scenarios

In some embodiments, the model allows the correlation between the effective age of a subject, the status of the subject’s body and his/her repair capacity. Referring now to Fig. 7 a graphical representation 700 of the difference in expected hypothesized healing behavior between young subjects and old subjects when faced with events and treatment is shown, according to some embodiments of the invention. The graph 700 shows a scale of health status over weeks. The full line 702 shows the expected behavior of a young subject’s recovery based of his/her health status. The line begins showing that the young subject feels well. Then at point 704 the subject falls ill. Since the subject is young, a relatively rapid recovery is expected. The long- dashed line 706 shows the expected behavior of an old subject’s recovery. Usually, since old people have accumulated damaging factors over the years, the beginning level of the health status is lower than that of a young subject. Thus, the line begins at some point between“well” and “weak”. At point 708 the old subject falls very ill. If the old subject would have a repair system ready and able to deal with the disease, the recovery graph would continue as depicted in rest of the long-dashed line 710. Since the repair resources available to the old subject are fewer compared to the young, the recovery time increases dramatically, as depicted by the short-dashed line 712. Another problem is that the available repair mechanism lessens at every event. For example, the old subject recovering from event 708 suffers another event at point 714. This second event, depicted as dotted line 716, brings the health status of the subject to worse than critical. The recovery behavior is expected to be long and slow due to the lack of repair mechanisms available to the old subject. The device and method of the present invention, provides an increase in the repair factors available to the subject, enhancing the repair and accelerating the recovery. If, for example, at point 718, the subject’s blood volume is increased following the device and methods of the present invention, the increased number of repair factors changes dramatically the recovery behavior, showed as the dashed-dotted line 720, potentially bringing the subject to a healthy status. In a more critical example, if at point 722 in dotted-line 716, the subject is treated with the present invention, the recovery behavior will improve, following the dashed-double dotted line 724, bringing the subject above the critical line in significantly shorter time. The improvement in repair behavior is much noted in critical cases. It will be obvious to any person having skill in the art, that providing the treatment of the present invention can help in a variety of cases, other than those mentioned above. For example, treatment to a subject of any age that has have suffered trauma, preventive treatment to any subject, treatment to athletes, and others.

As mentioned above, in some embodiments, the model allows interrelating between the effective age of a subject and his/her repair capacity. The repair capacity can be calculated as the number of senescent cells present in the subject. It has been shown that young subjects have more effective and numerous repair capacities (less senescent cells) than old subjects (more senescent cells). The senescent cells saturate the existing repair resources, and thus the more senescent cells, the fewer resources are free. Therefore, the number of senescence cells can be used to evaluate the "capacity to repair", in the model. Using this reference, in some embodiments, the efficacy of certain treatments are translated to“the decrease in number of senescent cells in relation to the expected (or actually measured) number of senescent cells in a subject”. For example, after treatment, the number of senescent cells, in an old subject, has decreased to a number similar to the number of senescent cells expected to be seen in a younger subject.

In some embodiments, sensitivity to model parameters is weak, since the increase in repair capacity entails a nearly proportional rejuvenation: Where: h represents the rate of increase in SC (senescent cells) production over time, b is the removal rate of SC, k is the halfway saturation point of SC clearance and e is the noise amplitude. Age is denoted by t. Mean senescent cell concentration is approximately (kpt+e)/(P-pt) which diverges at t-b/h (which occurs at old age) so an increase in maximal repair rate b causes a large decrease in mean senescence without specific sensitivity to other model parameters.

Exemplary relation between treatment and repair

Referring now to Fig. 8, showing in a series of graphs 800, the concept of how a treatment of increasing the total blood volume in a subject translates into a better repair capacity measured by the number of senescent cells in said subject, according to some embodiments of the invention. For example, as shown in the bottom left graph 810, doubling blood volume in an 80 years old subject (e.g.: a 4 to 6 liters increase in blood volume) will reduce senescent cell levels to the level expected to be seen in a 65 years old subject within one month and to the level expected to be seen in 55 years old subject within two months. For a subject of 60 years old, this will result in a reduction of senescent cell levels to that expected in a subject of 40 years old within one month and to that expected in a 35 years old subject within two months. This means a rejuvenation of two decades of major physiological processes. As shown in the middle left graph 806, a 50% blood volume increase in a subject of 80 years old will reduce damage levels similar to a 70 years old subject after two months. Lifespan is predicted to also increase by similar proportions. The change in the quantity of senescent cells can be seen as a "rejuvenation" result of the treatment, since the result is evaluated in relation to the number of senescent cells expected to be seen in a younger subject.

As can be seen in the graphs 800, in some embodiments, the benefits of the treatment are expected to persist for several months after cessation of treatment. For example, as can be seen in the bottom right graph 812, a 2-fold increase of blood volume in an 80 years old subject results in rejuvenation to a 65 years old subject after 30 days. Assuming that treatment stops after 30 days, cellular senescence level should be equivalent to that of a 72.5 years old subject, two months after cessation of treatment. Similar results can be seen for 1.25-fold increase in blood volume 804 and for 1.5-fold increase volume 808.

Referring now to Fig. 9, the rejuvenation (in terms of reduction in senescent cell numbers) of being connected to the device for different periods of time, as a function of age and blood volume enhancement, is shown 900. For example, as shown in graph 912, at day 45 having double relative blood volume, a rejuvenation from age 80 to 60 means that the number of senescent cells in the tissues reduces from its mean value at age 80 to a new value equal to the mean value of at age 60.

Referring now to Fig. 10, the rejuvenation after cessation of treatment (in terms of reduction in senescent cell numbers) of being connected to the device for different periods of time, as a function of age and blood volume enhancement, is shown 1000. For example, in graph 1012, at day 45 having double relative blood volume, a rejuvenation from age 80 to 70 means that the number of senescent cells in the tissues reduces from its mean value at age 80 to a new value equal to the mean value of at age 70.

Summarizing, in some embodiments, a certain increase in the blood volume for a certain period of time, shows:

Increase in the number of repair factors, which means a decrease in damage factors;

Decrease in the number of senescent cells, which means a decrease in damage factors.

Furthermore, in some embodiments, providing an increase in blood volume concomitantly to a drug that reduces damage factors provides a synergistic effect, strongly reducing damage factors in the body of a subject.

As mentioned above, in several embodiments of the present invention, the increase in blood volume is provided by a device. Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details of construction and the arrangement of the components and/or methods set forth in the following description and/or illustrated in the drawings and/or the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.

Exemplary embodiments

In some embodiments, the device is presented in different formats according to different requirements and/or constrains. Referring now to Figs l la-c, showing schematic representations of examples of some embodiments 1100 of the present invention are disclosed. One example of an embodiment is shown in Fig. l la, which is an external device 1102, attached to a subject 1104 while in bed 1106. Another example of an embodiment is shown in Fig. 1 lb, which is an external device 1120 carried by a subject 1122 optionally in a bag or any other carrying device 1124.

Another example of an embodiment is an internal device (implant) 1140 which is implanted inside the subject 1142.

Exemplary external generic embodiment

In some embodiments, the device is an external device. Referring now to Fig. 12, a schematic representation 1200 of an example of the embodiment of the external device 1102 is shown. In some embodiments the device is designed to be connected to a subject while he is in bed or has limited movement. In some embodiments, on one side, the device is connected to the circulatory system allowing the flow of blood to pass from the subject to the device. In some embodiments, the connection to the circulatory system is located at the internal jugular and femoral veins. In some embodiments, the connection to the circulatory system is located at the subclavian vein. Referring now to Fig. 13, a schematic representation of some embodiments 1300 of vascular access is shown. In some embodiments, the vascular access is performed via a central venous catheter (CVC) 1302. In some embodiments, implantable ports 1304 are used to facilitate connection and disconnection of the device. In some embodiments, the implantable ports help in case of unscheduled disconnection from the device. In some embodiments, to minimize the blood pressure originating from the heart, the device may be, for example, connected to a vein (e.g. to the inferior vena cava around the abdomen). In some embodiments, on the other side, the device is again connected to the circulatory system allowing the flow from the device to the circulatory system. In some embodiments, the connection of the device creates a bypass, rendering the extra volume of blood contained in the device available to the body.

In several embodiments, the device comprises means to add saline, blood, plasma, or blood substitutes (e.g. colloid) to allow volume expansion. In some embodiments, all blood component concentrations will return to homeostasis by physiological feedbacks (e.g. the innate immune cells (NK cells) that remove senescent cells recover after depletion on the timescale of one day. In some embodiments, red blood cells recover on a similar timescale. In some embodiments, the production of new blood cells by the subject’s bone marrow stems cells will need to increase (e.g. to double), which is thought to have no clinical consequence. In some embodiments, the device will require means to add EPO to ensure red blood cell volume homeostasis. In some embodiments, the blood from the body flows 1202 into the device via a tube 1204. In some embodiments the tube has a radius from about 3 millimeter (mm) to about 15 mm. In some embodiments the tube has a radius from about 4 mm to about 10 mm. In some embodiments the tube has a radius from about 5 mm to about 8 mm. In some embodiments, the device comprises means to supply or deliver anticoagulants 1206 to the entering blood from the body (e.g.: citrate, warfarin, heparin or CPDA). In some embodiments the device comprises means to measure the blood pressure 1208 entering the device. In some embodiments, these measurements are important since it is pivotal to keep the pressure’s homoeostatic levels of the body. In some embodiments, for minimal load on the circulatory and respiratory systems, blood flow through the device is minimal flow such that blood cells in the device remain unstressed, and immune cells in the blood do not exhaust. This is opposed to the higher flow rates desired for effective hemodialysis. For example, a flow rate of 6L/h=100ml/min.

In some embodiments, the contact time between NK cells and their targets are on the order of an hour, suggesting that a turnover time of ~l hour (e.g. lOOml/min) is sufficient for equal average cytotoxic activity of NK cells. Therefore, in some embodiments, the turnover time is from about 20 minutes to about 2 hours. In some embodiments, the turnover time is from about 40 minutes to about 1.5 hours. In some embodiments, the turnover time is from about 50 minutes to about 1.2 hours.

In some embodiments, the blood flow is maintained by gravity, by corporeal blood pressure, by compression of skeletal muscle or by an artificial pump 1210. In this kind of embodiment, where the subject is mainly in a horizontal position (lying down in a bed), the artificial pump 1210 has an important role. In some embodiments, the circulation is assisted by means of one-way valves in one or more locations in the tubes which can mimic vein skeletal muscle pump and pulmonary pumps. In some embodiments, the blood then flows into a container 1212 having a sizable volume of the subjects’ blood and/or other physiological liquids (e.g.: saline, artificial blood, etc.). In several embodiments of the invention, the device maintains viable blood volume with viable blood-bome repair factors in said container 1212. The material of the container 1212 may be biological such as synthetic organs or synthetic blood vessels, or non- biological containers, including glass, polycarbonate, polypropylene or polyethylene containers, or a combined material with a synthetic tube or matrix coated with endothelial cells. In some embodiments, the container’s materials are adapted to contain and maintain the user’s blood without damaging it. In some embodiments, bio-engineering of new blood vessels using bioprinting, stem cells or other cells expanded either in-vivo or ex-vivo can also provide the material.

In some embodiments, the pump works having a regular frequency. In some embodiments, the regular frequency is adapted to have the same frequency as the user’s heartbeat. In some embodiment, the pump works at a frequency other than a regular frequency. In some embodiments, the pump’s activation having a regular frequency same as the user’s heartbeat, comprises allowing incoming of flow into the device on a first beat, and allowing the outcoming of blood from the device on a second beat.

In some embodiments, the volume of the container is from about 0.5 liters to about 10 liters. In some embodiments, the volume of the container is from about 1 liter to about 8 liters. In some embodiments, the volume of the container is from about 3 liters to about 6 liters.

In some embodiments, the container 1212 may be a sack. In some embodiments, the container 1212 may be a syringe. In some embodiments, the container 1212 may be a long tube. In some embodiments, the container 1212 is a fixed-volume container. In some embodiments, the container 1212 is a volume-variable container. In these cases, the container comprises an adjusting (controlling) blood volume system, including means for reducing or increasing blood volume in case of need. In some embodiments, the enlargement of the volume of the container can be achieved by prolonging/shortening or expanding/narrowing the tube and/or the container, or by modifying the volume of the syringe, or by adding a series of valves that can be opened/closed, or by adding a buffer container of adjustable volume that can be adjusted where blood may be contained. Shortening the tube will decrease resistance, so it may be required to also narrow it to avoid increasing the velocity of the blood flow. In some embodiments, the control of the volume of the container is performed manually. In some embodiments, the control of the volume of the container is performed electronically by a dedicated controller.

In some embodiments the blood can be held in N tubes of length L and width a mm. In some embodiments, the resistance of the device is approximately R~8r_|L/(Nua^A4 ) where h is the viscosity of blood. In some embodiments, the volume of blood stored in the device is V=N a^A2 L. Where N is the number of tubes; a is the radius of each tube; L is the length of each tube; and h is the viscosity of blood. In some embodiments, assuming that the point of exit of blood from the body to the device is higher than the point of entry of blood to the device and the height difference is h, the pressure due to gravity is P=pgh where p is the density of the blood. In some embodiments, where the point of exit of blood from the body to the device is not positioned higher than the point of entry of blood to the device, a pump 1210 (i.e.: peristaltic pump) will be used. In some embodiments, a one-way valve is used and the movement of the blood is relied on thoracic breathing (pulmonary pump mechanism for venous flow).

An example of implementation is as follows: blood will flow through a single tube (N=l) of radius a=5mm and length L=30m (tube wound around itself). In some embodiments, tubes with radius exceeding 3mm are known to support long-term blood flow without clotting or obstruction. In some embodiments, the point of entry to the device is 5cm below the point of exit (h=5cm). In some embodiments, the tube will contain -2.5 liters of blood, and the flow rate through the tube will be 4.8 liters/hour. In some embodiments, when an individual is horizontal (e.g. during sleep) circulation will depend on muscle contraction or on an artificial pump. In some embodiments, one-way valves in the tube can help provide a skeletal muscle pump/pulmonary pump as in veins.

In some embodiments, from the container 1212, the blood continues to flow. In some embodiments, before leaving the device, the blood may be supplied with an antidote 1214 (e.g.: calcium) to neutralize the effect of the anticoagulant.

In some embodiments, the return flow from the device to the user is characterized by a continuous return flow, meaning: flow comes in the device, same flow exits the device. In some embodiments, the return flow from the device is limited to specific timings and quantities of flow, meaning: flow comes in, the device is full and ready to provide return flow to the body. Then, in some embodiments, at selected times, a selected amount of liquid is allowed to flow back to the circulatory system of the user. In some embodiments, the return flow is governed by “drip chamber”-1ike device, which receives liquids from the container in a drip-like manner, allowing the return flow to be regulated by the quantity of liquids entering the“drip chamber”- like device.

Another example of implementation for an implant: comprises a subcutaneous container 1212 (e.g.: similar to a large flattened infusion bag under skin around belly) having for example 2.5 liters (L), with a pump 1210, such that the energy supply is external to the body and connected by wires/induction to the pump 1210. In some embodiments, the pump 1210 will generate a flow of lOL/hour. In some embodiments, the container 1212 is connected to the vasculature using tubes 1204 and implanted CVC 1302 to the two femoral veins. In some embodiments, one of the main issues is clotting, which is resolved using a micro-pump that secretes anticoagulant 1206 (e.g.: citrate) at the entry port and an antidote 1214 (e.g.: calcium) at the exit port. In some embodiments, the device further comprises a de-aeration chamber 1216 to avoid the insertion of bubbles to the circulatory system. In some embodiments, after the de-aeration chamber 1216, an air-bubble detector 1218 is placed to ensure that no bubbles have passed. In some embodiments, the sensor may be coupled to a valve that shuts the flow in case bubbles are detected.

In some embodiments, at the end of the tube, a return pressure gauge 1220 controls the pressure of the blood that returns to body 1222.

In some embodiments, the device comprises means to abort the additional blood volume 1224 to return to normal volume circulation. In some embodiments, the abortion of additional blood is performed manually. In some embodiments, for ease of connection and abortion, a catheter may be used. In some embodiments, this point of exit can be used also as a point for entry to prime the device. In some embodiments, a separate point of entry is used for the insertion of liquids/drugs/other. In some embodiments, the device can be filled (primed) for example with

(i) the persons own blood (autologous blood) collected in the period before the procedure starts,

(ii) compatible donated whole blood or plasma or blood processed in any way (iii) the persons own blood from circulation. In some embodiments, the device will start with a small extra blood volume and will slowly increase this volume over time so that the subject generates the extra blood continuously (e.g. 5-10% of total volume/week). In some embodiments, the blood volume is monitored by using a standard tracer dilution technique (e.g.: 1-131 albumin injection). In some embodiments, the adjustments are done by increasing the container tube lengths or by having a series of containers that can be replaced. In some embodiments, after reaching the desired extracorporeal blood volume, the device continues to circulate the increased blood volume.

In several embodiments, the device comprises means for measuring and adjusting hemodynamics 1226, including means to measure and adjust the level or concentration of blood volume, arterial blood pressure, central venous pressure, oxygen delivery, cardiac output and pulmonary artery catheterization.

In some embodiments, arterial blood pressure is measured for example by using a sphygmomanometer 1228; central venous pressure is measured for example by an inserted catheter; blood oxygen levels are measured for example by pulse oximetry 1230; cardiac output is measured for example by a thermodilution method apparatus 1232; blood volume is measured for example by a tracer dilution technique 1234. In some embodiments, the means for measuring hemodynamics are integrated in the device. In some embodiments, the means for measuring hemodynamics are allocated outside the device.

In some embodiments, the device comprises means of oxygenation (e.g.: a pumpless oxygenator circuit) that is used when the blood flow rate is low. In some embodiments, the means of oxygenation are allocated near the exit part of the blood in the device. In some embodiments, the means for oxygenation are integrated in the device. In some embodiments, the means for oxygenation are allocated outside the device.

In some embodiments, the device comprises means for monitoring the correct functionality of the device 1236. In some embodiments, the correct functionality of the device is measured by an actual reduction in senescent cell count in the body. The reduction of senescent cells is measured by measuring: (i) senescent T-cells cells in blood; and/or (ii) NK cell function.

In some embodiments, the measurements are performed outside the device, using laboratory tests for e.g.: pl6 mRNA in T cells; external cytotoxic activity test for NK cells from blood using standard methods.

In some embodiments, the measurements are performed outside inside the device, using microfluidic versions of the assays in which blood cells are isolated into chambers, probed with the reagent (for pl6mRNA quantification by RT PCR on chip for example), and the results read by a microscopy system on-chip and processed using a computer; and/or microfluidics which isolate NK cells into chambers, and provide them with bait cells and measure their activity by cytotoxic assays.

In some embodiments, dedicated feedback sensors will monitor the correct functionality of the different parts of the device.

In some embodiments, the reduction in senescent cells is predicted to manifest in reduction of inflammation. Therefore, in several embodiments, inflammation and Senescence- Associated Secretory Phenotype (SASP), are externally monitored by:

Measuring and tracking levels of SASP associated cytokines, including IL-8, IL-6, IL-la, IL-Ib and other cytokines in blood or in tissues to detect SASP levels, by sandwich enzyme-linked immunosorbent assay or by other means, for example as described by Amsen et. al. in "Approaches to Determine Expression of Inflammatory Cytokines", Methods Mol Biol. 2009; 511: 107-142.

Measuring erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and plasma viscosity (PV) or other measurements to detect the level of inflammation by combining flow cytometry and image analysis, for example as described by Biran et. al. in " Quantitative identification of senescent cells in aging and disease", Aging Cell. 2017 Aug; 16(4): 661-671.

Monitoring senescent cells in blood, including Senescence-associated beta-galactosidase (SA-P-gal) assays and measurement of pl6Ink4A expression by quantification of pl6ink4a mma in T-cells, for example as described by Liu et. al. in "Expression of pl6INK4a in peripheral blood T-cells is a biomarker of human aging", Aging Cell. 2009 Aug; 8(4): 439-448, specifically for: senescent immune cells in blood, including measurements of senescence in circulating leukocytes.

It is expected that, in some embodiments, the device will improve cytotoxicity of immune cells and reduce exhaustion of immune cells. In some embodiments, to monitor this improvement, the device comprises:

Cytotoxicity assays for NK cells and other cytotoxic lymphocytes, for example as described by Sagiv et. al. in "NKG2D ligands mediate immunosurveillance of senescent cells", Aging (Albany NY). 2016 Feb; 8(2): 328-344;

Viability assays for target cells, such as assays for redox potential of the cell population, the integrity of cell membranes, or the activity of cellular enzymes such as esterases.

In some embodiments, the assays are performed in a micro-fluidic device, for example serial dilution micro-fluidic device.

In some embodiments, the monitoring data is stored on a dedicated server. In some embodiments, doctors and/or physicians may compare the output of the measurements to the expected output during the course of treatment and intervene accordingly. For example, if the device fails to reduce inflammation, other means may be chosen. In some embodiments, in case of an emergency, the doctor should either gradually reduce blood volume of the device, or, in extreme cases, remove the device entirely while keeping the patient under intensive care.

In some embodiments, the device further comprises means for adding drugs, nutrients, additives, reagents, and cells to blood. In some embodiments this can be done using port 1224. In some embodiments, a specialized device is attached to the device. This specialized device is able to monitor and provide the additives to the body via the device.

In several embodiments, the device comprises means (e.g.: thermostat) to maintain the temperature of the blood in the device to avoid cooling down or overheating of large extra corporeal quantities of blood.

In several embodiments, the device comprises an internal power source 1242 that provides the required power to all the parts that require energy. In several embodiments, the device comprises a controller 1244 which is adapted to monitor and control all the electronics of the device. In some embodiments, the device is further connected to a Graphic Unit Interface (GUI) 1246, which can be a specialized GUI provided with the device, or it can be a tablet or cell phone. In some embodiments, the GUI provides means to control and monitor the device; in some embodiments, it provides alerts to the subject in case there are problems; in some embodiments, it can stop the operation of the machine and can produce reports.

In some embodiments, the device operates following the generic method 1400 as described, for example, in Fig. 14. In some embodiments, the device is connected to the user 1402, then the valves are opened 1404. In some embodiments, at this point, monitoring the user is done 1406. In some embodiments, once the expected result is achieved, the valves are closed 1408, and, in some embodiments, the device is disconnected 1410. In some embodiments, once the expected result is achieved, the valves are closed 1408, but the device stays connected for further treatments 1412.

In some embodiments, the monitoring 1406 comprises several parallel processes, for example, as depicted in Fig. 15. In some embodiments, the monitoring methods 1500, begin by asserting that the valve is open 1502. If the valve is open, then the monitoring of the treatment and the device begin 1504. As mentioned above, in some embodiments, several monitoring processes are performed in parallel. Examples of monitoring are as follow: In some embodiments, if blood is coming in into the device 1506, then there is a release of anticoagulant 1508. In some embodiments, if blood is coming out the device 1510, then there is release of the anticoagulant’s antidote 1512. In some embodiments, if the income blood pressure is low 1514, then the pump is activated 1516. In some embodiments, if the outcome blood pressure is low 1518, then the pump is activated 1520. In some embodiments, if bubbles are found in the exiting blood 1522, then the exit valve is closed 1524. In some embodiments, other monitoring processes are performed during the activation of the device, for example: monitoring leakage, blood temperature, and more.

Exemplary portable external generic embodiment

In some embodiments, the device is a portable device that can be carried by the subject and is adapted to sustain the expected movement of the subject. In some embodiments, the device can be carried in a bag or in a belt attached to the body of the subject. Referring now to Fig. 16, an example of embodiment 1120 of the present invention is shown. In some embodiments, one of the main differences between this embodiment and the one disclosed before, is the configuration of the parts of the device. In some embodiments, the device need to be smaller and lighter than the one disclosed before.

In some embodiments, all the parts of the device are made of more robust materials, for example, the connectors must be more robust in order to avoid coagulation.

In some embodiments, the device includes the means (e.g. micro-pump) to provide the anticoagulant at the entry point and the antidote (e.g. also by a micro-pump) to neutralize the anticoagulant at the exit point 1602. In some embodiments, the device comprises means to measure the blood pressure entering the device; and the return pressure gauge 1604. In some embodiments, the device may include an artificial pump 1606 to maintain blood flow, or it may be maintained by gravity, by corporeal blood pressure or by compression of skeletal muscle.

In some embodiments, at the exit point the device comprises a de-aeration chamber 1608 to avoid the insertion of bubbles to the circulatory system. In some embodiments, after the de aeration chamber 1608, an air-bubble detector 1610 is placed to ensure that no bubbles have passed. In some embodiments, the sensor may be coupled to a valve that shuts the flow in case bubbles are detected.

In some embodiments, the device comprises several sensors 1612, as explained above, and a power source 1614. In some embodiments, the device comprises a GUI 1616 as well, similar to that disclosed above.

In some embodiments, this type of device, that is connected intermittently, requires means to prepare and store whole blood extra-corporeally for l2-24h or for several days (either in ambient temperature or cooling to 2-6 degrees Celsius). In some embodiments, the preparation of whole autologous blood entails adding an anticoagulant such as CPDA, and storing the blood in a temperature-controlled cabinet at the desired temperature. In some embodiments, room temperature is optimal temperature for immune cells in whole blood, which, in some embodiments, is the interesting fraction (e.g.: NK cells).

Exemplary internal generic embodiment

In some embodiments, the device is adapted to be implanted for long periods of time inside the body of the subject. Referring now to Fig. 17, an example of embodiment 1140, an internal device implanted in the body of the subject, is shown. In some embodiments, the internal device may require ventricular assist device such as a pump, use gravity, body muscle energy to move blood or use arterial pressure to move blood. In some embodiments, one-way valves in tubing can mimic vein skeletal muscle pump and pulmonary pumps. In some embodiments, the implant may be synthetic, organic or a combination of both. In some embodiments, the implant may be an extension of an artificial transplantable organ such as an artificial heart or artificial kidney, with increased volume for additional blood. In some embodiments, the implant may include artificial or surgically constructed blood vessels. In some embodiments, an implementation of the internal device is as follows: blood will flow through a single tube (N=l) of radius a=5mm and length L=30m, arranged spirally around itself. In some embodiments, tubes with radius exceeding 3mm support long-term blood flow without clotting or obstruction. In some embodiments, the point of entry to the device is 5cm below the point of exit (h=5cm). In some embodiments, the tube will contain -2.5 liters of blood, and the flow rate through the tube will be 4.8 liters/hour. In some embodiments, when an individual is horizontal (e.g. during sleep) circulation will depend on muscle contraction through a pulmonary pump mechanism. In some embodiments, one-way valves in the tube can help provide a skeletal muscle pump/pulmonary pump as in veins. In some embodiments, the preferable locations for vascular access are the internal jugular and femoral veins. In some embodiments, the subclavian vein is considered the third choice because higher risk of thrombosis. In some embodiments, vascular access is performed via a central venous catheter (CVC). In some embodiments, there is no interaction of the device with the external world regarding energy, fluid change, control, etc. In some embodiments, a pump can assist with the flow through the device, which may require wire to the external world to an extracorporeal energy supply (battery).

In several embodiments, the implant device comprises all the general means for device as described above.

Exemplary external embodiment

Referring now to Fig. 18, an example of an embodiment 1800 of the present invention is disclosed. In some embodiments, where the external devices or the internal implant devices described above cannot be used, the increase in blood volume is performed for example by increasing the vasculature of the subject, by using a specialized device 1802.

In some embodiments, the blood volume may be increased by one of or a combination of an induction of an arterio-venous shunt (fistula), surgically or by other means, or as a result of an implant functioning as a low-resistance circuit. Arteriovenous fistula increases blood volume by 0%-l0%.

In some embodiments, application of hormones that increase total blood volume are supplied, such as somatotropin or estrogen or other hormones that, for example, mimic the blood- volume enhancing effect of pregnancy. In some embodiments, estrogen causes a substantial increase in blood volume (a 20% increase in total blood volume due to estrogen in sheep). In some embodiments, the duration of the treatment is for at least three weeks. In some embodiments, the treatment is monitored as above (measurement of total blood volume, e.g.: by tracer dilution methods). In some embodiments, the treatment can be either transdermal (50pg/day) or oral (0.45 mg/day).

In some embodiments, application of means for vasodilation: one example can be pharmacological means for vasodilation (for example hydralazine, adenosine, or prostacyclin). Another example may be mechanical or electrical means, for example by applying repeated cycles of electrical stimulation.

In some embodiments, total blood volume is increased by induction of angiogenesis, either pharmacologically, such as by becaplermin, or by other means, such as by laser revascularization. In some embodiments, the application of laser revascularization is performed, for example, as described by Horvath et. al. in "Transmyocardial laser revascularization: Operative techniques and clinical results at two years" May l996Volume 111, Issue 5, Pages 1047-1053. In some embodiments, subsequent applications of laser revascularization to different blood vessels will take place until desired blood volume increase will be reached (monitored by tracer dilution). In some embodiments, becaplermin will be taken at a dosage of as per standard use for several weeks until desired blood volume increase will be reached.

In some embodiments, total blood volume is increased by pharmacological means to alter blood osmotic pressure, increase net fluid retention and decrease urine output, or other means that mimic the blood-volume enhancing effect of exercise, or drugs that interfere with epinephrine/norepinephrine release. These may include the following medications: Fludrocortisone (0.1 mg per day for several weeks); Erythropoietin (5000 IU for several weeks); Long term intake of clonidine Saline Solution (0.150 mg, several weeks); Vasopressin; Atenolol (Tenormin) (doses of 25/50/100 mg); Propanolol (Inderal) (doses of about 30 mg); Disopyramide (Norpace) (dosage: Immediate-release: 150 mg PO q6hr; Controlled-release: 300 mg ql2hr; Range: 400-800 mg/day); Angiotensin converting enzyme inhibitors/angiotensin receptor blockers (Depends on the drug, for example, for telmisartan the dosage will be 20mg/40mg/80mg for several weeks). In some embodiments, the duration of the treatment will continue until the desired blood volume is achieved, according, for example, to tracer dilution measurements.

In some embodiments, any implementation that should increase blood volume by up 50% can imitate the blood volume enhancement observed in pregnancy in the following manner: an induction of one or multiple arteriovenous shunts can imitate the blood volume enhancing effects of the placenta and may increase blood volume by 30%; applying estrogen therapy as mentioned above should increase blood volume by 20%.

In some embodiments, where applicable, the subject will be further subjected to at least one of the following, to increase the blood volume results in treated subjects:

In some embodiments, the treatment will be complemented by instructing the subject to perform exercise since it can also enhance blood volume by an additional 10%.

In some embodiments, the treatment will be complemented by treatments to increase immune surveillance (repair of senescent cells) in the body: for example, drugs that increase biogenesis of immune cells (NK cells, monocytes or T-cells) or raise their set-point in the blood.

In some embodiments, the treatment will be complemented by providing drugs that stimulate in-vivo expansion of immune cells such as IL-2 or IL-15.

In some embodiments, the treatment will be complemented by providing autologous or external (donor, cell culture, iPS or stem cell derived) supply of NK cells, monocytes or CD4+ T- cells.

In some embodiments, the treatment will be complemented by enhancing the activity of checkpoint inhibitors to increase NK-cell/T-cell cytotoxicity for senescence cells.

In some embodiments, the treatment will be complemented by providing drugs that improve homing/chemotaxis of NK-cells or T-cells to senescent cells.

In some embodiments, the treatment will be complemented by providing senolytic drugs - drug that directly target senescent cells.

In some embodiments, the treatment will be complemented by decreasing damage production rate by caloric restriction or by drugs that target mTOR pathway such as rapamycin or metformin.

In some embodiments, the device comprises an automatic shut-down mechanism which is activated in case the device is disconnected from the user at an unscheduled moment (e.g. disconnected by mistake, disconnected due to involuntary pulling of the device from the user.

Exemplary treatment criteria

In some embodiments, the physician chooses to treat the subject by connecting him to the device. In some embodiments, different criteria for treating the subject is as described, for example, in flowchart 1900 in Fig. 19. In some embodiments, the physician begins by checking the status of the subject 1902. In some embodiments, the physician will decide according to the status 1904 of the subject. In some embodiments, the criteria on which the physician choses to treat with the device is the level of frailty of the subject. In some embodiments, the criteria on which the physician choses to treat with the device is that the individual is considered frail 1906. In some embodiments, the criteria on which the physician choses to treat with the device is the level of trauma of the subject 1908. In some embodiments, the criteria on which the physician choses to treat with the device is the expectation of a procedure which may increase the possibilities of the subject to improve a frailty state 1912. In some embodiments, the criteria on which the physician choses to treat with the device is the frailty of the individual causes him to have high risk of mortality and causes a serious disability. In some embodiments, the criteria on which the physician choses to treat with the device is the individual is eligible for the treatment 1910: he or she have a functional immune system and are not at too high risk of complications from any procedure associated with the device. In some embodiments, the assessment of frailty is done following the procedures, for example, as described by Qian-Li Xue in "The Frailty Syndrome: Definition and Natural History" Clin Geriatr Med. 2011 February; 27(1): 1-15. In some embodiments, if the subject meets the criteria, then the physician will connect the subject to the device 1914. In some embodiments, if the subject does not meet the criteria 1916, then the physician will not connect the subject to the device 1920.

In some embodiments, the doctors may also decide to use the device to treat specific age- related conditions, such as osteoarthritis, diabetes, heart failure, kidney disease, dementia, neurodegenerative disease and any other age-related disease. In some embodiments, the device will help alleviate the frailty-related syndromes.

In some embodiments, the device may be used are seemingly non-frail individuals (including young individuals) that require extra repair capacity, for example after severe infection, injury or trauma. In some embodiments, the device will shorten the recovery time.

In some embodiments, the connection of the device to the user is fixed, and only can be removed by the assistance of specialized medical personnel. In some embodiments, the connection of the device to the user is dynamic, meaning, it can be connected and disconnected effortlessly. In some embodiments, the connection of the device to the user comprises a plug- in/plug-out mechanism, easily to operate by the user itself.

Exemplary criteria methods - Frailty

Frailty is theoretically defined as a clinically recognizable state of increased vulnerability resulting from aging-associated decline in reserve and function across multiple physiologic systems such that the ability to cope with every day or acute stressors is comprised.

This definition of frailty (multisystem dysregulation yielding decreased physiologic reserves and increased vulnerability to stressors) has commonality to that of aging (loss of molecular/cellular functional properties yielding decreased adaptability to intemal/extemal stress and increased vulnerability to disease and mortality). Both have a basis in loss of homeostasis, although with aging, the failure in homeodynamics is global, whereas with frailty, the failure in homeodynamics cycle around energy metabolism and neuromuscular changes (Fedarko N.S., The Biology of Aging and Frailty, Clin Geriatr Med 27 (2011) 27-37).

In some embodiments, criteria for treatment is evaluated by the frailty state of the patient. In some embodiments, the frailty state is assessed using the Linda Fried / Johns Hopkins Frailty Criteria and/or the Rockwood Frailty Index and/or the Four domains of frailty model and/or the SHARE Frailty Index and/or any other frailty criteria that will be created in the future.

Exemplary computerized diagnostic-therapy tool

In some embodiments, the model is inserted into a computer program adapted to evaluate several parameters related to the patient, the repair status of the patient, the damage status of the patient, the current health state of the patient, and provide possible treatment methods using the device disclosed herein, optionally with the addition of pharmacological treatment.

In some embodiments, the computer program will receive continuous updates from the device and/or the physician and/or the patient, and amend or stop the chosen treatment. In some embodiments, the computer program will provide updates to the physician on the state of the patient, and will provide suggestions according to the model and to collected data over time.

In some embodiments, a dedicated server is used to collect data from the different devices and/or physicians and/or patients and create an anonymous database which will collect all information regarding patients, health states, results of treatments, etc. This database is used to improve the model over time.

In some embodiments, any of the abovementioned treatments are performed for as long as it is needed until the desired effect is reached and/or the desired blood volume is achieved. In some embodiments, the treatments are performed following an intermittent regimen (e.g.: 1 month on, 3 months off). In some embodiments, the treatment can be performed in only part of the day, for example only during the night.

Exemplary blood incubation

As disclosed above, senescent cells are a major cause of aging-related decline and morbidity. In some embodiments, blood from a subject is taken and incubated for a period of time outside the body of the subject. In some embodiments, incubation of the blood outside the body causes the restoration of the removal function of NK cells, thereby enhancing the removal of senescent cells from the corporeal blood once the blood is returned to the body. In some embodiments, the incubation period is from about 6 hours to about 24 hours. Optionally, the incubation period is from about 8 hours to about 36 hours. Optionally, the incubation period is longer than 36 hours. In some embodiments, using the device as described above, the blood is taken from the subject, but instead of returning it right away, the blood is left outside the body, incubating for a period of time. In some embodiments, while extracting the blood from the subject, new blood is inserted into the body of the subject to replace the extracted blood. Optionally, the new blood is blood after incubation period. In some embodiments, the extracted blood is incubated in the device while still attached to the subject. In some embodiments, the device comprises a plurality of compartments where blood is incubated and, once the incubation period is over, the device is activated and the blood is returned to the subject. In some embodiments, the blood replacement/incubated treatment is performed for from about one week to about 4 weeks. Optionally, for from about two weeks to about 6 weeks. Optionally for from about 4 weeks to about 8 weeks. In some embodiments, daily checks of senescent cells level in the blood are measured to monitor the efficacy of the treatment. In some embodiments, optionally monitoring is performed every couple of days. Optionally once a week. A potential advantage of this process is to provide an alternative to drugs, based on extracorporeal blood volume and autologous blood transfusions with reactivated NK cells.

As used herein with reference to quantity or value, the term“about” means“within ± 20% of’.

The terms“comprises”,“comprising”,“includes”,“including”,“has”,“having” and their conjugates mean“including but not limited to”.

The term“consisting of’ means“including and limited to”.

The term“consisting essentially of’ means that the composition, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.

As used herein, the singular forms“a”,“an” and“the” include plural references unless the context clearly dictates otherwise. For example, the term“a compound” or“at least one compound” may include a plurality of compounds, including mixtures thereof.

Throughout this application, embodiments of this invention may be presented with reference to a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as“from 1 to 6” should be considered to have specifically disclosed subranges such as“from 1 to 3”,“from 1 to 4”,“from 1 to 5”,“from 2 to 4”,“from 2 to 6”,“from 3 to 6”, etc.; as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

Whenever a numerical range is indicated herein (for example“10-15”,“10 to 15”, or any pair of numbers linked by these another such range indication), it is meant to include any number (fractional or integral) within the indicated range limits, including the range limits, unless the context clearly dictates otherwise. The phrases“range/ranging/ranges between” a first indicate number and a second indicate number and“range/ranging/ranges from” a first indicate number“to”,“up to”,“until” or“through” (or another such range-indicating term) a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numbers therebetween.

Unless otherwise indicated, numbers used herein and any number ranges based thereon are approximations within the accuracy of reasonable measurement and rounding errors as understood by persons skilled in the art

As used herein the term“method” refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

As used herein, the term“treating” includes abrogating, substantially inhibiting, slowing or reversing the progression of a condition, substantially ameliorating clinical or aesthetical symptoms of a condition or substantially preventing the appearance of clinical or aesthetical symptoms of a condition.

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements. Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

In addition, any priority document(s) of this application is/are hereby incorporated herein by reference in its/their entirety.

Claims

WHAT IS CLAIMED IS:

1. A method for increasing blood volume in a body of a subject, comprising:

a. attaching a container with a volume of at least 25% of the volume of the circulatory system of said subject to the circulatory system of said subject;

b. allowing the flow of blood from said body to said container; and

c. allowing the flow from said container back to said body;

thereby increasing the blood volume of said circulatory system of said subject.

2. The method according to claim 1, wherein at the initial moment of attaching, said container is filled with at least one liquid other than blood.

3. The method according to any one of claims 1-2, wherein said attaching is from outside said body.

4. The method according to any one of claims 1-3, wherein said attaching is from inside said body.

5. The method according to any one of claims 1-4, wherein said allowing the flow comprises controlling said flow by a valve.

6. The method according to any one of claims 1-5, wherein said method further comprises providing said subject with at least one drug.

7. The method according to claim 6, wherein said drug is selected from the group consisting of: senolytic drug, hormones, vasodilatation drugs, pro-angiogenic drugs, antibiotics, steroids, anticoagulants, and any combination thereof.

8. The method according to any one of claims 1-7, wherein said method further comprises monitoring homeostatic parameters in said subject’s circulatory system.

9. The method according to any one of claims 1-8, wherein said allowing the flow comprises allowing the flow to flow in a continuous manner.

10. The method according to any one of claims 1-9, wherein said allowing the flow comprises allowing the flow to flow in an intermittent manner.

11. The method according to any one of claims 1-10, wherein said attaching is performed for a period of time of at least one week.

12. A device for increasing blood volume in the body of a subject, comprising:

a. at least one first tube having a first extremity and a second extremity; said first extremity interconnected in continuous flow to the circulatory system of a subject and said second extremity to at least one container;

b. at least one container operatively interconnected to said second extremity of said at least one first tube; said at least one container adapted to contain at least one liquid;

c. at least one second tube having a third extremity and a fourth extremity; said third extremity operatively interconnected to said container; said fourth extremity interconnected in continuous flow to said circulatory system of said subject;

characterized in that the flow from said circulatory system through said container back to said circulatory system is a low flow;

characterized in that said flow from said circulatory system through said container back to said circulatory system does not change the homeostatic state of said circulatory system; and characterized in that said at least one container is adapted for safely containing the blood of said subject.

13. The device according to claim 12, further comprising means to deliver at least one anticoagulant into said at least one first tube.

14. The device according to claim 13, further comprising means to deliver at least one antidote to said anticoagulant into said at least one second tube.

15. The device according to any one of claims 12-14, further comprising means to monitor said homeostatic state of said circulatory system inside said device.

16. The device according to any one of claims 12-15, further comprising at least one sensor.

17. The device according to claim 16, wherein said sensor is selected from the group consisting of: blood pressure, bubble sensor, arterial blood sensor, blood coagulation sensor, pH sensor, central venous sensor, oxygen sensor, cardiac output sensor, temperature sensor, and any combination thereof.

18. The device according to any one of claims 12-17, further comprising a de-aeration chamber.

19. The device according to any one of claims 12-18, further comprising a two-way port into said container; said two-way port interconnected to at least one third tube.

20. The device according to any one of claims 12-19, further comprising means of oxygenation of said at least one liquid.

21. The device according to any one of claims 12-20, further comprising means for monitoring the correct functionality of said device.

22. The device according to any one of claims 12-21, further comprising means for controlling the temperature of said at least one liquid.

23. The device according to any one of claims 12-22, further comprising at least one power source.

24. The device according to any one of claims 12-23, further comprising at least one controller.

25. The device according to any one of claims 12-24, further comprising at least one graphic unit interface.

26. The device according to any one of claims 12-25, wherein said device is an external device.

27. The device according to any one of claims 12-25, wherein said device is an internal device.

28. The device according to any one of claims 12-27, wherein said device is a portable device.

29. A method of choosing a treatment for a subject, comprising:

ii. receiving said frailty score according to a frailty assessment model; b. choosing a treatment according to said frailty score.

30. The method according to claim 29, wherein when said frailty score is lower than a predetermined frailty score value, then providing pre-treatment to increase said frailty score above said predetermined frailty score value.

31. The method according to claim 29, wherein said frailty assessment model is based on at least one model selected from the group consisting of:

a. Linda Fried / Johns Hopkins Frailty Criteria model;

b. Rockwood Frailty Index model;

c. Four domains of frailty model;

d. SHARE Frailty Index model; and

e. any combination thereof.

32. A method of affecting a frailty status of a subject according to a frailty score assessment tool, comprising:

ii. receiving said frailty score according to a frailty assessment model; b. when said frailty score is lower than a predetermined frailty score value, then treat with device according to claim 12.