System and method for extracorporeal blood treatment

Abstract

The present application relates to an extracorporeal blood treatment system (50) for treating a subject, the system comprising: an extracorporeal blood circuit (52); a dialysate fluid circuit (54); the extracorporeal blood circuit and the dialysate fluid circuit being separated by a membrane (56) of a filtration unit (58); at least one blood pump (60) for controlling the flow of blood through the blood circuit; at least one dialysate fluid pump (62) for controlling the flow of dialysate fluid through the dialysate fluid circuit (54); a system computing unit (64) operatively connected to the blood pump and to the dialysate fluid pump, the system computing unit having at least one input; wherein the system computing unit is adapted to receive a desired blood concentration of glutamine GLN b , to receive a desired blood concentration of glucose GLUCOSE b , and to receive a desired blood concentration of ketone bodies KETONE b ; the system computing unit being adapted to control the blood pump and the dialysate fluid pump such that an actual concentration value of glutamine GLN a is driven to approach GLN b , and an actual concentration value of glucose GLUCOSE a is driven to approach D b , and an actual concentration value of ketone bodies is driven to approach KETONE a . The present application relates to a treatment device for use in an extracorporeal blood treatment system, a method for treating cancer using the system, a controller for controlling the method and a dialysis liquid suitable for use in the method.

Description

Technical Field

[0001] The present disclosure relates herein to extracorporeal blood treatment. More specifically, the present disclosure relates to the use of extracorporeal blood treatment, such as hemodialysis, in treating cancer, as well as systems for such treatment and user interfaces for controlling and displaying data recorded during such extracorporeal blood treatment (sg historical data). Background Art

[0002] Most human cancer cells show an altered energy metabolism that distinguishes them from normal cells. Normal cells obtain most of their energy through mitochondrial oxidative phosphorylation, the aerobic process of oxidizing glucose, first through glycolysis and then through the tricarboxylic acid (TCA) cycle to produce adenosine triphosphate. On the contrary, this pathway is secondary only in cancer cells. This was first observed by Warburg in the 1920s. He noted that after cancer cells have metabolized glucose through glycolysis, lactate is produced from pyruvate. In normal cells, this only occurs under anaerobic conditions, but in cancer cells, even in the presence of a large amount of oxygen, this alternative pathway is increased. This phenomenon is referred to as "aerobic glycolysis" or "Warburg effect" (Warburg et al., 1927, Gen Physiol 8:519-530). Subsequent studies have confirmed the presence of a characteristic glycolytic phenotype in cancer cells, which has also been observed to overexpress enzymes related to glycolysis in most cancer cells.

[0003] The above metabolic transformations give cancer cells a selective growth advantage and contribute to their ability to resist hypoxia and apoptosis. Because the rate of tumor cell proliferation exceeds the rate of new blood vessel formation, many tumors grow in low-oxygen environments. There are various metabolic changes in cancer cells, and the most common and well-known is their habit of producing energy through aerobic glycolysis. Moreover, many intermediates of glycolysis (such as, for example, ribose, glycerol and serine) are also intermediates of biosynthetic pathways and anabolic pathways necessary during cancer cell growth and proliferation. In addition, glycolysis produces ATP from ADP, which allows to maintain cell growth in tumors. However, glycolysis is much less efficient than oxidative phosphorylation, and therefore requires a large amount of glucose to produce sufficient amounts of ATP. Therefore, this metabolic pathway requires a large amount of glucose. Many cancer cells are addicted to glucose as their main energy supplier. For a variety of reasons, if their glucose supply is targeted, glycolytic tumor cells become vulnerable.

[0004] Furthermore, many cancer cells also display an addiction to glutamine. Glutamine-dependent cells exhibit high rates of glutamine uptake not only due to its role as a nitrogen source in nucleotide and amino acid biosynthesis, but also because glutamine is a major mitochondrial matrix in cancer and is required for the production of NADPH for redox control and macromolecular synthesis.

[0005] In cancer, there are other metabolic changes that play an important role in survival, and importantly, many cancers show a surprisingly good ability to change their metabolic profile. This plasticity to withstand environmental challenges (such as when glucose, glutamine or oxygen become low) is important for the survival of cancer cells.

[0006] Furthermore, many cancer cells also display a dependence on glutamine. The high rates of glutamine uptake exhibited by glutamine-dependent cancer cells are due not only to its role as a nitrogen source for nucleotide and amino acid biosynthesis, but also to the fact that glutamine is the major mitochondrial metabolic substrate in many cancers and is required for the production of NADPH for redox control and macromolecular synthesis.

[0007] In cancer, there are many metabolic alterations, and various amino acids such as glutamine have important roles in cancer metabolism to control redox balance and generate building blocks for continued proliferation. Moreover, when new metabolic constraints need to be imposed, many cancers show a surprisingly good ability to utilize those alternative pathways to change their metabolic map. This quality (ability) is an important feature of cancer and its ability to withstand environmental challenges when metabolic energy resources such as glucose, glutamine or oxygen become scarce.

[0008] Therefore, in order to effectively affect cancer through metabolic approaches, it is important to influence its metabolic system from more than one direction. Reduced glucose can be easily processed by most cancer cells, but if several metabolic pathways are affected simultaneously (such as reducing glucose and glutamine), the sum of these changes becomes more damaging than the individual parts.

[0009] In addition to glucose, ketones, and glutamine, which serve as global energy sources for many cancers, serine and glycine meet important specific requirements for maintaining cell growth and proliferation in cancer, for example, through one-carbon metabolism. In addition to their extensive energy requirements, cancer cells must also accumulate building blocks for the construction of new cellular components (including nucleic acids, proteins, and lipids), as well as equally important cofactors for maintaining their cellular redox state (Amelio et al.: Trends. Biochem. Sci. (2014), Vol. 39(4): 191-198).

[0010] Studies have shown that arginine is essential for cell growth and can become limiting in rapidly growing states and also affect survival if removed from cancer cells (Albaugh et al., J. Surg. Oncol. (2017), Vol. 115(3), 273-280).

[0011] In view of the above, it has been proposed that lowered blood sugar can be used as a strategy for a wide range of glycolysis-dependent tumors. Under hypoglycemic conditions, fat, especially ketone bodies, can replace glucose as the main metabolic fuel for normal cells. However, many tumors have abnormalities in the genes and enzymes required to metabolize lipids and ketone bodies for energy. Therefore, the conversion of energy from carbohydrates to ketones specifically points to energy metabolism in glycolysis-dependent tumor cells (Seyfried et al., 2010, Nutrition and Metabolism, 7:7).

[0012] According to this approach, for example, WO2011070527 discloses a method for treating a proliferative disease (cancerous or non-cancerous) in an individual, wherein a hemodialysis machine is used to lower blood glucose concentration.

[0013] Using a hemodialysis machine to lower blood glucose has the following advantages: it can reduce the glucose concentration in the blood, thereby reducing it in a more controlled and effective manner compared to dietary glucose deprivation. However, the method and apparatus disclosed in WO2011070527 require blood glucose sensors and blood glutamine sensors connected to the blood inlet flow, blood return flow, and dialysate, all of which are connected to the central control unit of the hemodialysis machine. In addition, the central control unit of WO2011070527 is also connected to an electroencephalogram (EEG) to provide information related to spontaneous electrical activity to the central unit to initiate an increase in glucose and glutamine levels. Such a large number of sensors and instruments leads to a high level of complexity and the associated high costs. In addition, patients receiving this treatment must consume a glucose-restricted diet for several days before treatment. This is not an insignificant burden for patients.

[0014] There is a constant need for improved methods of treating cancer. Summary of the Invention

[0015] In a first aspect of the present invention, there is provided an extracorporeal blood processing system (50) for treating a subject suffering from cancer, the system comprising:

[0016] Extracorporeal blood circuit (52a, 52b);

[0017] a dialysate fluid circuit (54a, 54b);

[0018] The extracorporeal blood circuit (52a, 52b) and the dialysate fluid circuit (54a, 54b) are separated by a membrane (59) of a filtration unit (58);

[0019] at least one blood pump (60) for controlling the flow of blood through the blood circuit (52a, 52b);

[0020] at least one dialysate fluid pump (62, 68) for controlling the flow of dialysate fluid through the dialysate fluid circuit (54a, 54b);

[0021] Optionally, one or more infusion lines (66, 80, 81, 82), each infusion line being connected to an extracorporeal blood circuit (52a, 52b) or adapted to be connected directly to the vascular system of a subject to be treated, each infusion line comprising an infusion pump;

[0022] a system computing unit (64) operatively connected to the blood pump (60) and the dialysate fluid pump (62, 68), and optionally to one or more infusion pumps of one or more infusion lines (66, 80, 81, 82), the system computing unit having a user interface comprising an input device and a display device;

[0023] The system calculation unit (64) is adapted to receive (901) a desired blood concentration value GLN of glutamine in the range of 0.1 mM and 0.5 mM b ;

[0024] The system calculation unit (64) is adapted to receive (903) a desired blood concentration value of glucose GLUCOSE in the range of 2 mM to 4 mM b ;

[0025] The system computing unit (64) is adapted to receive (905) a desired blood concentration value of a ketone body such as acetoacetate, β-hydroxybutyrate or a pharmaceutically acceptable derivative, ester or salt of acetoacetate or β-hydroxybutyrate in the range of 1 mM to 15 mM KETONE b ;

[0026] The system computing unit (64) is adapted to receive (902) a concentration value GLN p , the concentration value GLN p represents the concentration of glutamine or a pharmaceutically acceptable glutamine-containing compound in fresh dialysate fluid;

[0027] The system computing unit (64) is adapted to receive (904) the concentration value GLUCOSE p , the concentration value of GLUCOSE prepresents the concentration of glucose in fresh dialysate fluid;

[0028] Optionally, the system computing unit (64) is adapted to receive (907) the concentration value KETONE p,, , the concentration value KETONE p,, represents the concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of acetoacetate and β-hydroxybutyrate in fresh dialysate fluid;

[0029] Optionally, the system computing unit (64) is adapted to receive (906) a concentration value KETONE via one of the one or more infusion lines (66, 80, 81, 82). i , the concentration value KETONE i represents the concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of acetoacetate and β-hydroxybutyrate in the infusion fluid to be infused into the extracorporeal blood line (52b) or directly into the vascular system of the subject to be treated;

[0030] The system calculation unit (64) is adapted to receive (911) the actual concentration value GLN of glutamine in the blood of the treated subject. a , and adapted to receive (910) an actual value of the concentration of glucose GLUCOSE in the blood of the treated subject a , and adapted to receive (911) actual concentration values ​​of ketone bodies such as acetoacetate and / or beta-hydroxybutyrate KETONE a ;

[0031] The system computing unit (64) is adapted to control the blood pump (60) and the dialysate fluid pumps (62, 68) such that the actual blood concentration value GLN of glutamine is a Driven (912) close to or below GLN b , and the actual blood concentration of glucose GLUCOSE a Driven (914) close to or below GLUCOSE b ;

[0032] and,

[0033] If the system (50) comprises one or more of the infusion lines (66, 80, 81, 82), and in case one of the infusion lines (66, 80, 81, 82) is adapted to infuse the infusion liquid into an extracorporeal blood line (52b) or directly into the vascular system of a subject to be treated, the system computing unit (64) is adapted to control the infusion pump of the infusion line such that the actual blood concentration value KETONE a Driven (920) close to KETONE b ;

[0034] or,

[0035] If the system (50) does not include such an infusion line (66, 80, 81, 82), the system computing unit (64) is adapted to convert the KETONE a with KETONE b Compare and if KETONE a <KETONE b , it is suitable to display a message on the display device to inform the subject of treatment that he should consume more ketone bodies or medium-chain triglycerides.

[0036] In the present disclosure, the term "subject" relates to a human or animal patient in need of treatment.

[0037] In a preferred embodiment, the extracorporeal blood treatment system (50) comprises one or more of said infusion lines (60, 80, 81, 82), and the system computing unit (64) is adapted to receive (906) a concentration value KETONE via one of said one or more infusion lines (66, 80, 81, 82). i , the concentration value KETONE i represents the concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of acetoacetate and β-hydroxybutyrate in the infusion fluid to be infused into the extracorporeal blood line (52b) or directly into the vascular system of the subject to be treated, and

[0038] The system computing unit (64) is adapted to control the infusion pump of the infusion line so that the actual blood concentration value KETONE a Driven (920) close to KETONE b .

[0039] In a preferred embodiment, the system computing unit (64) is adapted to monitor GLN a and GLUCOSE a , and if GLN a Lower than GLNb , it is suitable to start (916) infusion of a composition comprising glutamine or a pharmaceutically acceptable compound comprising glutamine by activating an associated infusion pump of one of the one or more infusion lines (66, 80, 81, 82), and / or, if GLUCOSE a Lower than GLUCOSE b , then is adapted to start (918) infusion of a composition comprising glucose by activating an associated infusion pump in one of the one or more infusion lines (66, 80, 81, 82), and is adapted to maintain the infusion until GLN a Equal to GLN b And GLUCOSE a Equal to GLUCOSE b .

[0040] The term "pharmaceutically acceptable glutamine-containing compound" refers to an oligopeptide, typically a dipeptide in which at least one of the amino acid residues is glutamine. Typical examples of such dipeptides are L-alanyl-L-glutamine and L-glycyl-L-glutamine. Glutamine-containing compounds are often used in place of glutamine in liquid compositions to enhance stability and solubility.

[0041] The term "ketone bodies" relates to water-soluble molecules containing a ketone group that can be produced by the liver from fatty acids. Typically, the ketone bodies according to the invention are β-hydroxybutyrate or a pharmaceutically acceptable derivative of β-hydroxybutyrate, such as its enantiomers (R)-β-hydroxybutyric acid, (S)-β-hydroxybutyrate or a mixture of enantiomers, or a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable ester thereof, and acetoacetate. According to the invention, medium-chain triglycerides are also considered to be derivatives of ketone bodies. The term "medium-chain triglycerides" or "MCT oil" refers to triglycerides with two or three fatty acids, with an aliphatic tail of 6 to 12 carbon atoms. Such medium-chain triglycerides or MCT oils can be converted into ketone bodies in the human body. Examples of infusion solutions containing ketone bodies or ketone body derivatives are MCT / LCT 20% (B.Braun) or 20% (Fresenius Kabi). Further examples can be found in WO2018 / 114309A1. Only the concentration of β-hydroxybutyrate and / or acetoacetate is detected in the blood of the subject as the actual ketone body concentration KETONE a , because other ketone body derivatives are converted into any of these compounds in the subject's body.

[0042] Preferably, the filtration membrane has a molecular weight cut-off (MWCO) of less than 60 kDa

[0043] More preferably, the filtration membrane has a MWCO of less than about 50 kDa or less than about 40 kDa, such as less than 30 kDa, less than 10 kDa, less than 5 kDa, or less than 2 kDa.

[0044] Preferably, the blood circuit comprises a thermal management system for heating or cooling the blood in the blood line during use.

[0045] More preferably, the thermal management system is controllable to adjust the temperature of the blood in the blood circuit to a temperature between 20°C and 43°C.

[0046] Preferably, the system (50) further comprises one or more sensors (S, 90, 91, 92, 93) for detecting an analyte selected from the group consisting of glucose, glutamine and ketone bodies, the sensors (S, 90, 91, 92, 93) preferably being positioned in the effluent portion (54a) of the dialysate fluid circuit, the sensors being in communication with the system computing unit (64) and providing an output indicative of the concentration of the analyte in the blood or preferably in the spent dialysate fluid; wherein the system computing unit (64) is configured to determine a representative blood concentration of the analyte from the output of the sensors (S, 90, 91, 92, 93) to thereby monitor GLN a 、GLUCOSE a and KETONE a At least one of .

[0047] In a second aspect, the present invention provides a treatment device for use in an extracorporeal blood treatment system (50) according to the first aspect, the device comprising a filter unit (58) having a membrane (59) which separates an integrated blood line (52a, 52b) and an integrated dialysate fluid line (54a, 54b), wherein the blood line (52a, 52b) and / or the dialysate fluid line (54a, 54b) comprises a sensor (S, 90, 91, 92, 93) for monitoring GLN a 、GLUCOSE a and KETONE a At least one of .

[0048] As shown below, suitable sensors and assays for analyzing glutamine, glucose, and ketone bodies are known in the art.

[0049] In a third aspect, the present invention provides a method of treating a subject having cancer using a system (50), the method comprising:

[0050] Extracorporeal blood circuit (52a, 52b);

[0051] a dialysate fluid circuit (54a, 54b);

[0052] The extracorporeal blood circuit (52a, 52b) and the dialysate fluid circuit (54a, 54b) are separated by a membrane (59) of a filtration unit (58);

[0053] at least one blood pump (60) for controlling the flow of blood through the blood circuit (52a, 52b);

[0054] at least one dialysate fluid pump (62, 68) for controlling the flow of dialysate fluid through the dialysate fluid circuit (54a, 54b);

[0055] and optionally, one or more infusion lines (66, 80, 81, 82), each infusion line being connected to an extracorporeal circuit (52a, 52b) or adapted to be connected directly to the vascular system of a subject to be treated, each infusion line comprising an infusion pump;

[0056] The method includes:

[0057] Receive (901) concentration value GLN b , the concentration value GLN b represents the expected blood concentration of glutamine in the range of 0.1 mM and 0.5 mM;

[0058] Receive (903) concentration value GLUCOSE b , the concentration value of GLUCOSE b represents the expected blood concentration of glucose in the range of 2 mM to 4 mM;

[0059] Receive (905) concentration value KETONE b , the concentration value KETONE b represents the desired blood concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate, or pharmaceutically acceptable derivatives, esters, and salts of β-hydroxybutyrate in the range of 1 mM to 15 mM;

[0060] Receive (902) the concentration value GLN p , the concentration value GLN p represents the concentration of glutamine or a pharmaceutically acceptable glutamine-containing compound in fresh dialysate fluid;

[0061] Receive (904) the concentration value GLUCOSE p , the concentration value of GLUCOSE p represents the concentration of glucose in fresh dialysate fluid;

[0062] Optionally, the system computing unit,

[0063] Optionally, receiving (907) a concentration value KETONE p,, , the concentration value KETONE p,, represents the concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of β-hydroxybutyrate in fresh dialysate fluid;

[0064] Optionally, a concentration value KETONE is received (906) via one of the one or more infusion lines (66, 80, 81, 82). i , the concentration value KETONE i represents the concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts thereof in the infusion fluid to be infused into the extracorporeal blood line (52b) or directly into the vascular system of the subject to be treated;

[0065] Receive (909) the concentration value GLN a , the concentration value GLN a represents the actual concentration of glutamine in the blood of the treated subject;

[0066] Receive (910) the concentration value GLUCOSE a , the concentration value of GLUCOSE a represents the actual concentration of glucose in the blood of the treated subject;

[0067] Receive (911) concentration value KETONE a , the concentration value KETONE a Indicates the actual concentration of ketone bodies such as acetoacetate and β-hydroxybutyrate;

[0068] The blood pump (60) and the at least one dialysate fluid pump (62, 68) are controlled so that the actual concentration value GLN of glutamine is a Driven (912) close to or below GLN b , and the actual concentration of glucose GLUCOSE a Driven (914) close to or below GLUCOSE b ;

[0069] and,

[0070] If the system (50) comprises one or more of the infusion lines (66, 80, 81, 82), and one of the infusion lines (66, 80, 81, 82) infuses the infusion liquid into the extracorporeal blood line (52b) or directly into the vascular system of the subject to be treated, the infusion is controlled so that KETONE a Driven (920) close to KETONE b ,

[0071] or,

[0072] If the system (50) does not infuse any infusion fluid containing ketone bodies, then the KETONE a Lower than KETONE b In the case of ketones, the subject to be treated is required to orally take a certain amount of ketone bodies or medium-chain triglycerides.

[0073] Preferably, the system (50) comprises one or more of the infusion lines (66, 80, 81, 82), and the method further comprises:

[0074] Receive (906) concentration value KETONE i , the concentration value KETONE i Indicates the concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of β-hydroxybutyrate in the infusion fluid;

[0075] infusing the infusion liquid into an extracorporeal blood line (52b) through one of the one or more infusion lines (66, 80, 81, 82) or directly into the vascular system of the subject to be treated; and

[0076] Control the infusion pump of the infusion line so that the actual blood concentration value KETONE a Driven (920) close to KETONE b .

[0077] Preferably, the cancer is selected from human colon cancer and glioblastoma, and prostate cancer, breast cancer and liver cancer.

[0078] In a fourth aspect, the present invention provides a system computing unit (64) adapted to control an extracorporeal blood processing system (50) for treating a subject suffering from cancer; the system computing unit comprising:

[0079] a plurality of output devices adapted to be operatively connected to at least one blood pump (60), at least one dialysate fluid pump (62, 68), and optionally one or more infusion pumps for controlling flow in each of the one or more infusion lines (66, 80, 81, 82);

[0080] a user interface comprising an input device and a display device; and

[0081] Memory devices and computing devices;

[0082] The system computing unit (64) is adapted to receive (901) a desired blood concentration value GLN of glutamine b ;

[0083] The system calculation unit (64) is adapted to receive (903) a desired blood concentration value of glucose GLUCOSE b ;

[0084] The system calculation unit (64) is adapted to receive (905) a desired blood concentration value of ketone bodies KETONE b ;

[0085] The system computing unit (64) is adapted to receive (902) a dialysate concentration value GLN of glutamine p ;

[0086] The system calculation unit (64) is adapted to receive (904) a dialysate concentration value of glucose GLUCOSE p ;

[0087] Optionally, the system computing unit (64) is adapted to receive (907) the concentration value KETONE p,, , the concentration value KETONE p,, represents the concentration of ketone bodies such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of β-hydroxybutyrate in fresh dialysate fluid;

[0088] Optionally, the system computing unit (64) is adapted to receive (906) a ketone body infusion fluid concentration value KETONE i ;

[0089] The system calculation unit (64) is adapted to receive (909) the actual blood concentration value GLN of glutamine a ;

[0090] The system calculation unit (64) is adapted to receive (910) the actual blood concentration value of glucose GLUCOSE a ;

[0091] The system calculation unit (64) is adapted to receive (911) an actual blood concentration value KETONE of a ketone body selected from the group consisting of beta-hydroxybutyrate and acetoacetate. a ;

[0092] The system computing unit (64) is adapted to control the blood pump (60) and the dialysate fluid pumps (62, 68) such that the actual blood concentration value GLN of glutamine is a Driven (912) close to or below GLN b , and the actual blood concentration of glucose GLUCOSE a Driven (914) close to or below GLUCOSE b ;

[0093] and,

[0094] In case the infusion pump is operatively connected to the system computing unit (64), the system computing unit (64) is adapted to control the infusion pump so that the actual blood concentration value KETONE a Driven (920) close to KETONE b ;

[0095] and,

[0096] In the absence of an infusion pump operatively connected to the system computing unit (64), the system computing unit (64) is adapted to convert the KETONE a with KETONE b Compare and if KETONE a <KETONE b , it is suitable to display a message on the display device to inform the subject of treatment that he should consume more ketone bodies or medium-chain triglycerides.

[0097] In a fifth aspect, the present invention provides a dialysis fluid suitable for dialysis treatment of cancer comprising a ketone body such as acetoacetate, β-hydroxybutyrate or a pharmaceutically acceptable derivative, ester or salt of β-hydroxybutyrate.

[0098] Preferably, the dialysis fluid further comprises a) glutamine or a compound comprising glutamine; and b) glucose.

[0099] Preferably,

[0100] a) the concentration of glutamine or a compound comprising glutamine is in the amount of 0 mM to 0.5 mM, preferably in the amount of 0.05 mM to 0.3 mM;

[0101] b) the concentration of glucose is in the amount of 0 mM to 6 mM, preferably in the amount of 0.5 mM to 4 mM; and

[0102] c) The concentration of ketone bodies is in the range of 1 mM to 15 mM, preferably in the range of 2 mM to 12 mM.

[0103] The above summary of the present disclosure is not intended to describe every embodiment or every implementation thereof. Advantages of the present disclosure and a more complete understanding of the present disclosure will become apparent and readily understood by referring to the following detailed description and claims in conjunction with the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0104] Figure 1 is a schematic diagram of a system according to the present invention when in use.

[0105] Figure 2 is a block diagram of an exemplary extracorporeal blood processing system including an input device and a display device that may employ the user interface and methods described herein.

[0106] Figure 3 is a perspective illustration of an exemplary dialysis system that may include a graphical user interface as described herein.

[0107] Figure 4 yes Figure 2 A front view of a portion of an exemplary dialysis system is shown.

[0108] Figure 5 is a schematic diagram of a portion of a system with thermal management according to an embodiment of the present invention.

[0109] Figures 6a to 6k Graph showing the results of Study 1.

[0110] Figures 7a to 7d Graph showing the results of Study 2.

[0111] Figure 8A 、 Figure 8B and Figure 8C The therapeutic objectives according to the present invention are shown.

[0112] Figure 9 is a flow chart illustrating the control algorithm of the present invention.

[0113] Figure 10 Shown are the growth rates of A549 cells and RCC4 cells in Medium A to Medium C as indicated.

[0114] Figure 11 The amounts of lung cancer A549 cells, renal cancer RCC4 cells, and primary RCC cells after culturing for 3 days in medium A to medium C under 21% or 5% oxygen are presented. Asterisks indicate significantly different values ​​determined by two-way ANOVA with Tukey's multiple comparison test.

[0115] Figure 12 Shown are the results of culturing human glioma cell line A172 under normoxia and hypoxia in medium A supplemented with 8 mM Acac, 16 mM BOHB, or a combination of 4 mM Acac / 8 mM BOHB to medium C. 4 mM LiCl and 8 mM LiCl were used as controls for Acac. Significantly different values ​​determined by one-way ANOVA using a Sidak multiple comparison test are marked by asterisks.

[0116] Figure 13 The results of culturing the glioma cell line U118MG under normoxia and hypoxia in medium A supplemented with 8 mM Acac, 16 mM BOHB, or a combination of 4 mM Acac / 8 mM BOHB are shown. 4 mM and 8 mM LiCl were used as controls for Acac. Significantly different values ​​determined by one-way ANOVA using a Sidak multiple comparison test are marked with asterisks.

[0117] Figure 14 Results of culturing the glioma cell line A172 under normoxia and hypoxia in medium B supplemented with 8 mM Acac, 16 mM BOHB, or a combination of 4 mM Acac and 8 mM BOHB to medium C are presented. 4 mM and 8 mM LiCl were used as controls for 4 mM Acac / 8 mM BOHB or 8 mM Acac, respectively. Significantly different values ​​determined by one-way ANOVA using a Sidak multiple comparison test are marked by asterisks.

[0118] Figure 15 Results of culturing the glioma cell line U118MG under normoxia and hypoxia in medium B supplemented with 8 mM Acac, 16 mM BOHB, or a combination of 4 mM Acac and 8 mM BOHB to medium C are presented. 4 mM and 8 mM LiCl were used as controls for 4 mM Acac / 8 mM BOHB or 8 mM Acac, respectively. Significantly different values ​​determined by one-way ANOVA using a Sidak multiple comparison test are marked by asterisks.

[0119] Figure 16 Results of culturing the renal cancer cell line RCC4 under normoxia and hypoxia in medium B supplemented with 8 mM Acac, 16 mM BOHB, or a combination of 4 mM Acac and 8 mM BOHB to medium C are presented. 4 mM and 8 mM LiCl were used as controls for 4 mM Acac / 8 mM BOHB or 8 mM Acac, respectively. Significantly different values ​​determined by one-way ANOVA using a Sidak multiple comparison test are marked by asterisks. DETAILED DESCRIPTION

[0120] Will refer to Figures 1 to 5 Describe the example system and method for selecting, viewing and filtering the historical data for extracorporeal blood treatment. Extracorporeal blood treatment system can record or store one or more parameters and / or events of one or more extracorporeal blood treatments, and generate historical data. The example system and method described in this article provide a graphical user interface, to display such historical data. Typically, historical data can include patient fluid removal data, fluid data, treatment data, anticoagulation data, pressure data, event data, setting data, patient data, alarm data, system voltage and current data, system timing data, user interaction data (e.g., interaction with user interface, such as button pressing and screen selection) etc. In the present invention, data can include the concentration of certain substances in the patient's bloodstream, because the system can include one or more sensors for detecting those concentrations. The substances detected can include glucose, glutamine and other amino acids (e.g., serine, glycine and arginine), ketones and cytokines.

[0121] Patient fluid removal data may include total patient fluid removal data, unplanned patient fluid removal data, selected limit data (e.g., selected limits for unplanned patient fluid gain / loss over a selected time period such as 1 hour, 3 hours, or 24 hours), etc. Fluid data may include pre-infusion data, dialysate data, post-replacement fluid filtration data, filtration data, effluent data, filtration fraction data, pre-dilution data, patient per kilogram rate data, ultrafiltration rate data, blood flow rate % post-ultrafiltration rate data, etc. Treatment data may include prescribed effluent dose data, delivered effluent dose data, target effluent dose data, prescribed ultrafiltration rate (UFR) dose data, target UFR dose data, delivered UFR dose data, etc. Anticoagulation data may include heparin data, estimated patient citrate load data, citrate solution data, calcium solution data, replacement solution data, calcium compensation data, syringe volume delivery data, bolus delivery data, etc. Pressure data may include inlet line pressure data, return line pressure data, filtration pressure data, transmembrane pressure (TMP) data, pressure drop across the filter (P DROP) data (e.g., pressure status in the blood compartment of the filter), self-test data, pressure alarm data, disconnection and occlusion limit data, stable pressure data, etc. Event data may include system configuration data, alarm data, setting data, treatment setting data, advisory data, prescription setting data, system setting data, anticoagulation data, pressure data, patient data, mechanical data, dosage data, etc.

[0122] Figure 1The depicted example extracorporeal blood processing system 10 can be used to perform the example methods and / or processes described herein. In at least one embodiment, the system 10 can be or include a machine for extracorporeal blood processing. The system 10 can alternatively be or include, for example, a blood processing device or a blood component preparation device or other medical device for fluid delivery and / or collection.

[0123] As shown, the example extracorporeal blood treatment system 10 includes a computing device 12. The computing device 12 can be configured to receive input from an input device 20 and send output to a display device 22. In addition, the computing device 12 can include a data storage device 14. The data storage device 14 can allow access to processing programs or routines 16 and one or more other types of data 18, which can be used to implement example methods and / or processes for use in performing extracorporeal blood treatment, recording historical data, filtering historical data, and displaying historical data. For example, the computing device 12 can be configured to record or log data such as flow rate and volume, to allow a user to select and view various sets of historical data using the input device 20 (e.g., based on input from the user), and to display the user-selected historical data using the display device 22.

[0124] The computing device 12 can be operably coupled to the input device 20 and the display device 22, for example, to transmit data to and from each of the input device 20 and the display device 22. For example, the computing device 12 can be electrically coupled to each of the input device 20 and the display device 22 using, for example, an analog electrical connection, a digital electrical connection, a wireless connection, a bus-based connection, etc. As further described herein, a user can provide input to the input device 20 to manipulate or modify one or more graphical depictions (e.g., windows, regions, areas, buttons, icons, etc.) displayed on the display device 22 to select and / or display historical data. In addition, various devices and apparatuses can be operably coupled to the computing device 12 for use within the computing device 12 to perform one or more in vitro procedures / treatments and the functions, methods, and / or logic described herein. As shown, the system 10 can include the input device 20 and the display device 22. The input device 20 can include any device capable of providing input to the computing device 12 to perform the functions, methods, and / or logic described herein. For example, the input device 20 may include a touch screen (e.g., a capacitive touch screen, a resistive touch screen, a multi-touch touch screen, etc.), a mouse, a keyboard, a trackball, etc. The input device 20 may allow a user to select and filter various historical data to be viewed on the display device 22 (e.g., display a graphical user interface depicting the historical data).

[0125] Likewise, the display device 22 may include any device capable of displaying information to a user (such as a graphical user interface, etc.) to perform the functions, methods, and / or logic described herein. For example, the display device 22 may include a liquid crystal display, an organic light emitting diode screen, a touch screen, a cathode ray tube display, etc. In at least one embodiment, a touch screen device may be placed on the display screen, allowing the user to touch graphical buttons and icons on the display to cause specific actions to occur.

[0126] As further described herein, the display device 22 can be configured to display a graphical user interface that includes one or more regions and / or areas for selecting and displaying real-time data and / or historical data for the extracorporeal blood treatment. For example, the graphical user interface displayed by the display device 22 can include or display a two-dimensional graph, a data set plotted on the two-dimensional graph, one or more graphical elements or icons representing events proximate to the two-dimensional graph, a time interval selection area, an event type selection area, an event list area or view, an event information area, a historical data area, an event display area, and the like.

[0127] A user may use each graphic, region, view, button, icon, panel, area, dialog, etc. to select and view historical data on the graphical user interface of the display device 22. As used herein, a "region" of a graphical user interface may be defined as a portion of a graphical user interface that may display information or perform a function. Regions may exist within other regions, may be displayed separately or simultaneously, etc. For example, a smaller region may be positioned within a larger region, regions may be positioned in parallel, etc. Additionally, as used herein, a "region" of a graphical user interface may be defined as a portion of a graphical user interface that is located in an area smaller than the region in which it resides.

[0128] The processing programs or routines 16 may include programs or routines for performing computational mathematics, matrix mathematics, normalization algorithms, comparison algorithms, or any other processing required to implement one or more of the example methods and / or processes described herein. The data 18 may include, for example, historical data, user accounts, licensing information, treatment profiles, bitmaps, videos, calibration data, system configuration information, solution data, engineering logs, event and alarm data, system pressure, system voltage, system current, self-test sequence data, user interaction data, process status data, monitor usage data, utilization data, software executables, patient information, process summary information, process runtime data, graphics (e.g., graphical elements, icons, buttons, windows, dialogs, drop-down menus, graphic areas, graphic regions, 3D graphics, etc.), graphical user interfaces, results from one or more processing programs or routines employed in accordance with the disclosure herein, or any other data that may be necessary to implement one and / or more of the processes or methods described herein.

[0129] In one or more embodiments, the system 10 can be implemented using one or more computer programs executed on a programmable computer (such as a computer including, for example, processing power, data storage (e.g., volatile or non-volatile memory and / or storage elements), input devices, and output devices). The program code and / or logic described herein can be applied to input data to perform the functions described herein and to generate desired output information. The output information can be applied as input to one or more other devices and / or methods as described herein, or can be applied in a known manner.

[0130] Any programmable language (e.g., a high-level procedural and / or object-oriented programming language suitable for communicating with a computer system) can be used to provide a program for implementing the methods and / or processes described herein. Any such program can, for example, be stored on any suitable device (e.g., a storage medium) that can be read by a general-purpose or special-purpose program running on a computer system (e.g., including a processing device) for configuring and operating the computer system when reading a suitable device for performing the processes described herein. In other words, in at least one embodiment, system 10 can be implemented using a computer-readable storage medium configured with a computer program, wherein the storage medium configured in this manner causes the computer to operate in a specific and predefined manner to perform the functions described herein. In addition, in at least one embodiment, system 10 can be described as being implemented by logic (e.g., object code) encoded in one or more non-transitory media including code for execution, and when executed by a processor, can be operated to perform operations such as the methods, processes, and / or functions described herein.

[0131] Likewise, system 10 may be configured at a remote site (e.g., an application server) that allows access by one or more users via a remote computer device (e.g., via a web browser) and allows the users to employ functionality according to the present disclosure (e.g., the users access a graphical user interface associated with one or more programs to process data).

[0132] The computing device 12 may be, for example, any fixed or mobile computer system (e.g., a controller, a microcontroller, a personal computer, a minicomputer, etc.). The exact configuration of the computing device 12 is not limiting, and essentially any device that can provide suitable computing power and control capabilities (e.g., graphics processing, control of extracorporeal blood treatment equipment, etc.) may be used.

[0133] As described herein, a digital file may be any medium (e.g., volatile or non-volatile memory, CD-ROM, punch cards, recordable tape, etc.) containing digital bits (e.g., encoded in binary, ternary, etc.) that may be readable and / or writable by a computing device 12 described herein.

[0134] Furthermore, as described herein, a file in a user-readable format may be any representation of data (e.g., ASCII text, binary numbers, hexadecimal numbers, decimal numbers, graphics, etc.) that can be presented on any medium (e.g., paper, a display, etc.) that can be read and / or understood by a user.

[0135] In view of the above, it will be apparent that the functions described in one or more embodiments according to the present disclosure can be implemented in any manner known to those skilled in the art. Likewise, computer languages, computer systems, or any other software / hardware used to implement the processes described herein should not limit the scope of the systems, processes, or programs described herein (e.g., the functions provided by such systems, processes, or programs).

[0136] It will be appreciated that a graphical user interface may be used in conjunction with the embodiments described herein. The user interface may provide various features that allow user input thereto, changes to input, the import or export of files, or generally any other features that may be suitable for use with the processes described herein. For example, the user interface may allow a user to select and filter various historical data to be displayed on a display device.

[0137] The methods and / or logic described in this disclosure, including those attributed to the systems or various components, may be implemented at least in part in the form of hardware, software, firmware, or any combination thereof. For example, various aspects of the technology may be implemented within one or more processors (including one or more microprocessors, DSPs, ASICs, FPGAs, or any other equivalent integrated or discrete logic circuits, as well as combinations of such components or other devices). The term "processor" or "processing circuitry" may generally refer to any of the foregoing logic circuits, alone or in combination with other logic circuits, or any other equivalent circuits.

[0138] Such hardware, software and / or firmware can be implemented in the same device or in separate devices to support the various operations and functions described in this disclosure. In addition, any of the components described can be implemented together or individually as discrete but interoperable logic devices. Depicting different features (e.g., using block diagrams, etc.) is intended to highlight different functional aspects and does not necessarily imply that such features must be implemented by separate hardware or software components. On the contrary, functions can be performed by separate hardware or software components, or integrated in common or separate hardware or software components. When implemented in software, the functions attributed to the systems, devices and methods described in this disclosure can be embodied as instructions and / or logic on a computer-readable medium (such as RAM, ROM, NVRAM, EEPROM, flash memory, magnetic data storage medium or optical data storage medium, etc.). Instructions and / or logic can be executed by one or more processors to support one or more aspects of the functions described in this disclosure.

[0139] The exemplary systems described herein for selecting and viewing historical data in extracorporeal blood treatments and the exemplary methods performed or used by such exemplary systems may generally be referred to as dialysis systems. The general term dialysis, as used herein, includes hemodialysis, hemofiltration, and hemodiafiltration. In dialysis, blood is typically removed from the body and exposed to a treatment device to separate substances from it and / or add substances to it, and then the blood is returned to the body. Therefore, reference herein to Figures 2 to 4 The example extracorporeal blood treatment system of describes an extracorporeal blood treatment system capable of performing general dialysis. Other systems may benefit from the systems, methods, and apparatus described herein, and the present disclosure is not limited to any particular fluid treatment system.

[0140] exist Figure 2 In the schematic diagram of FIG, an example extracorporeal blood treatment system 50 generally includes a blood circuit 52, which includes an arterial line 52a and a venous line 52b, which can be connected to the patient's vascular system. The device includes a filtration unit 58, which has a main chamber (blood chamber) and a secondary chamber (dialysis fluid chamber) separated by a semipermeable filter membrane 59. The inlet of the main chamber is connected to the blood withdrawal line or arterial line 52a. Similarly, the outlet of the main chamber is connected to the blood return line or venous line 52b.

[0141] The inlet of the secondary chamber is connected to a fresh dialysate fluid supply line 54b, which in turn is connected to a source 74 for providing fresh dialysis solution. The outlet of the secondary chamber of the filtration unit 58 is connected to a dialysis solution or spent dialysate line 54a, which delivers spent dialysis solution to an effluent connector 73.

[0142] In the present invention, filtration unit 58 includes a membrane 59 that differs from conventional hemodiafiltration membranes in that it is configured to remove relatively small compounds from the blood in blood line 112. The membrane is configured to have a molecular weight cutoff (MWCO) of 50 kDa or less, or even 40 kDa or less, such as 30 kDa or less, 10 kDa or less, 5 kDa or less, or 2 kDa or less.

[0143] The flow of blood through the blood lines 52a, 52b and the filtration unit 58 is controlled by a blood pump 60 positioned in the arterial line 52a. Similarly, the flow of fresh dialysate fluid from the source 74 through the filtration unit 58 to the effluent collector 73 is controlled by the dialysate fluid pump 68 in the supply line 54b, while the pressure in the dialysate fluid line 54 is controlled by a pressure relief valve in the spent dialysate line 54a. Alternatively, in place of the pressure relief valve, a second pump 62 may be included in the spent dialysate line 54a.

[0144] There may also be one or more infusion lines 66, 80, 81, 82 connected to the venous portion of the blood line 52b. In some embodiments, one or more of these infusion lines are adapted to be directly connected to the patient's vascular system (not shown). Each of the one or more infusion lines 66, 80, 81, 82 may include a separate pump. In one embodiment, an infusion fluid containing ketone bodies or ketone body derivatives may be infused through such infusion lines.

[0145] The system computing unit 64 is configured to communicate with the blood pump 60, the dialysate fluid pump 68, and one or more pumps of one or more infusion lines 66, 80, 81, 82, as well as a pressure relief valve or additional dialysate pump 68, so as to provide control of those devices during use. In an alternative embodiment, the system computing unit 64 provides appropriate setting data to the user via a graphical user interface for the pumps in one or more infusion lines 66, 80, 81, 82. The system computing unit is also in communication with an input device for providing information and instructions to the system computing unit 64. The input device may include a graphical user interface, such as a graphical user interface that can be controlled using a touch screen arrangement. In some embodiments, the input device may include a keyboard. In some embodiments, the input device may include one or more sensors for detecting the concentration of various substances or states in the patient's blood and / or spent dialysate.

[0146] For example, sensors may be provided to detect the concentration of one or more of amino acids (such as glutamine, serine, glycine, arginine), glucose, ketones, and cytokines in the subject's blood. In particular, such sensors should be suitable for detecting the concentration of one or more of glutamine, glucose, and ketone bodies in the subject's blood. Such sensors S, 91, 93 may be provided in the blood line 52 or using a separate device S, 90 that may be placed elsewhere on the patient's body. To the extent they are deployed, the sensors provide an indication of the actual blood concentration of the desired analyte to the system computing unit 64 (directly or via an additional input device). This provision may be performed periodically or substantially continuously during treatment.

[0147] Suitable sensors and assays for analyzing glutamine, glucose, and ketone bodies are known in the art. For example, a glutamine sensor is disclosed in US Pat. No. 4,780,191. Examples of glutamine assays are disclosed in US Pat. No. 9,995,750 and US Pat. No. 2016 / 168619. Examples of sensors and assays for determining ketone body concentrations are disclosed in US Pat. No. 8,532,731 B2, WO Pat. No. 2016 / 178823, US Pat. No. 5,326,697, and US Pat. No. 5,618,686. Examples of sensors and assays for glucose are disclosed in US Pat. No. 2019 / 328,288, US Pat. No. 2018 / 128,767, and US Pat. No. 2011 / 105,871.

[0148] Furthermore, the patient's blood concentrations of amino acids (e.g., glutamine, serine, glycine, and arginine) and one or more of glucose, ketones, and cytokines (preferably, at least glucose and glutamine) can be measured by drawing and separately analyzing (e.g., laboratory analysis) the patient's blood. Although the facility can be deployed during any treatment period, depending on the proximity and speed of the analysis, this method may be particularly relevant during prolonged treatment.

[0149] In another embodiment, one or more sensors S, 92 may optionally be included in the effluent line 54b. The sensor may detect the concentration of one or more of glucose, amino acids (such as glutamine, serine, glycine, and arginine), ketones, or cytokines, as desired. The concentration of the relevant analyte measured in the effluent line may be used to determine a representative concentration of the relevant analyte in the patient's blood, such as in the manner described in European Patent No. 2377563 (which is incorporated herein by reference).

[0150] When cells in the body are exposed to temperatures higher than normal, changes occur within the cells. Depending on the extent of the area being treated, the treatment can be local (tumor only), regional (as in a limb), or whole-body hyperthermia. Very high temperatures can cause necrosis / apoptosis (thermal ablation) of cancer cells, but high temperatures can also damage or induce apoptosis / necrosis of normal cells. Therefore, hyperthermia must be carefully controlled.

[0151] Hyperthermia is a promising method for improving cancer treatment, but today it requires specialized equipment and is therefore cumbersome to perform. Many clinical trials for the treatment of cancer with hyperthermia are underway. The proposed additional physical circuit is well suited to induce and control hypothermia in patients. Raising blood temperature while treating extracorporeal blood content forms a good combination therapy that can enhance the effect of altered blood content.

[0152] At a blood flow of 300 mL / min and a temperature of 43 degrees in the returning blood, an effect of approximately 120 watts is delivered to the patient, and the body temperature will easily rise. The blood temperature can be raised to 45°C in a short time without substantially damaging the blood.

[0153] The patient may need to be cooled to terminate the treatment, but it is also necessary to regulate the treatment that is too effective. Cooling will result in a reduction in the metabolic rate of normal cells and cancer cells, but will subsequently also result in a reduction in their oxygen consumption. The changes mentioned will result in protecting normal healthy cells, but will result in the weakening of cancer cells to, for example, conventional cancer treatments, and will also result in cancer dialysis and the reduction of glucose and glutamine and oxygen. Cooling can also result in less adverse side effects of the conventional cancer treatment given to the patient, and will also withstand the more active treatment of cancer dialysis and conventional cancer treatments. This accompanying drawing does not indicate any device for cooling or heating blood, but this mechanism is well known to those skilled in the art, and is often used in combination with some types of extracorporeal blood treatments, such as continuous renal replacement therapy (CRRT).

[0154] Figure 8A 、 Figure 8B and Figure 8C The therapeutic objectives and principles of the present invention are illustrated.Each figure discloses an example of normal conditions with respect to the concentration of a specific blood component. Figure 8A It is shown that patients typically have actual blood concentrations of glutamine GLN in the range of 0.20 mM to 0.8 mM at the start of treatment a During the treatment period, the actual blood concentration of glutamine was reduced to the desired value GLN b , the desired value is in the range of 0.1 mM to 0.5 mM, and for example, in the range of 0.15 mM to 0.3 mM. Figure 8BIt shows that patients typically have actual blood concentrations of glucose in the range of 4 mmol / l to 8 mmol / l at the start of treatment. a During the treatment, the actual blood concentration of glucose is reduced to the desired value GLUCOSE b , the expected value is usually in the range of 2mmol / l to 4mmol / l. Finally, Figure 8C It is shown that the blood initially contains almost no ketone bodies (such as β-hydroxybutyrate or a physiologically acceptable salt or ester of β-hydroxybutyrate (such as the sodium salt). Therefore, the actual value of the ketone body concentration in the patient's blood ketones is about 0. Usually, fresh dialysate does not contain any ketone bodies or contains only a small amount of ketone bodies. During treatment, by infusing solutions of ketone bodies and / or ketone body derivatives, the blood concentration of such ketones can be increased to the desired value in the range of 1 mmol / L to 15 mmol / L KETONE a (such as the desired value KETONE in the range of 2mM to 12mM b ).

[0155] In one embodiment, the actual blood concentration of glutamine GLN a and / or actual blood glucose concentration GLUCOSE a Can be lower than the expected GLN b and / or GLUCOSE b These actual blood concentration values ​​GLN can then be adjusted by infusing a solution containing glucose and / or glutamine or a pharmaceutically acceptable glutamine-containing compound. a and / or GLUCOSE a Increase to the expected value GLN b and / or GLUCOSE b .

[0156] Figure 9 A flow chart of an example of an algorithm 900 for a control process according to the present invention is disclosed. Before starting treatment, the system computing unit 64 is adapted to receive treatment target concentrations or desired blood concentrations of key compounds. Typically, these treatment target concentrations are input using a user interface.

[0157] Therefore, in step 901, the system computing unit 64 receives the desired blood concentration value GLN of glutamine. b .

[0158] In step 903, the system computing unit 64 receives the expected blood glucose concentration value GLUCOSE b .

[0159] In step 905, the system computing unit 64 receives the expected blood concentration value of ketone bodies KETONEb .

[0160] Before starting the treatment, the system computing unit 64 is also adapted to receive concentration values ​​for the above key components in the fresh dialysate fluid. Typically, these concentration values ​​are also input using a user interface.

[0161] Therefore, in step 902, the system computing unit 64 receives the concentration value GLN of glutamine in the fresh dialysate fluid. p .

[0162] In step 904, the system computing unit 64 receives the glucose concentration value GLUCOSE in the fresh dialysate fluid. p .

[0163] In step 907, the system computing unit (64) is adapted to receive the concentration value KETONE p,, , the concentration value KETONE p,, Indicates the concentration of ketone bodies (such as acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of β-hydroxybutyrate) in fresh dialysate fluid;

[0164] In step 906, the system computing unit 64 receives the ketone body concentration value KETONE in the infusion fluid to be infused in the extracorporeal blood line (52b) or directly into the vascular system of the subject / patient to be treated. i .

[0165] Before starting the treatment, and optionally during the treatment, the system computing unit 64 is adapted to receive actual concentration values ​​determined from the patient's blood. These concentration values ​​can be obtained after collecting a blood sample and subsequently analyzing it in a separate analysis unit. The concentration values ​​are then manually entered via the user interface. In some embodiments, the system computing unit is connected to one or more of these separate analysis units and can therefore receive data directly from them. In some embodiments, the blood circuits 52a, 52b and / or the dialysate circuits 54a, 54b may include suitable sensors S, 90, 91, 92, 93 that are connected to the system computing unit 64 receiving the data. The system computing unit 64 typically calculates the actual concentration values ​​GLN based on the most recently received actual concentration values ​​GLN. a 、GLUCOSE a and KETONE a To control the processing.

[0166] Therefore, in step 908, the system computing unit 64 begins processing.

[0167] In step 909, the system computing unit 64 is adapted to receive the actual blood concentration value GLN of glutamine from the patient. a .

[0168] In step 910, the system computing unit 64 is adapted to receive the actual blood concentration value of glucose GLUCOSE from the patient. a .

[0169] In step 911, the system computing unit 64 is adapted to receive the actual blood concentration value of ketone bodies KETONE from the patient. a .

[0170] In step 912, the system computing unit 64 controls the blood pump 60 and the dialysate fluid pumps 62, 68 so that GLN a Driven close to or below GLN b .

[0171] In a simultaneous step 914, the system computing unit 64 controls the blood pump 60 and the dialysate fluid pumps 62, 68, so that GLUCOSE a Driven close to or below GLUCOSE b .

[0172] In a simultaneous step 916, the system computing unit 64 monitors the GLN a , and if GLN a Lower than GLN b , the system computing unit starts and maintains infusing the composition containing glutamine or a compound containing glutamine into the extracorporeal blood circuit 52a, 52b.

[0173] In a simultaneous step 918, the system computing unit 64 monitors GLUCOSE a , and if GLUCOSE a Lower than GLUCOSE b , the system computing unit starts and keeps infusing the glucose-containing composition into the extracorporeal blood circuits 52a, 52b.

[0174] In a simultaneous step 920, the system computing unit 64 monitors K a , and if K a Lower than K b , the system computing unit starts and maintains the infusion of the composition containing ketone bodies into the extracorporeal blood circuit 52a, 52b or directly into the vascular system of the subject to be treated.

[0175] In some embodiments, the concentrations of other substances in the blood can be controlled. For example, it has been found that lowering the concentrations of amino acids such as serine, glycine, and arginine from normal is beneficial.

[0176] exist Figures 3 and 41 and 12. The diagrams in the perspective view and partial front view of an example extracorporeal blood treatment system 110 in which treatments and graphical user interfaces as described herein may be implemented generally include a blood circuit 112 having a first tubing segment 114 and a second tubing segment 116 connected to the vascular system of a patient 118 via an inlet device 117 and a return device 119, respectively. As will be understood by those skilled in the art, devices 117 and 119 may be cannulas, catheters, or winged needles, among others. Tube segments 114 and 116 are further connected to a filtration or treatment unit 120. In dialysis, filtration unit 120 is a dialyzer, also commonly referred to as a filter.

[0177] Many other component devices of the blood circuit 112 are also included, such as, for example, pressure sensors 127, 128, 154, 129. Additional sensors may be provided to monitor the concentrations of glucose and / or glutamine, ketones (such as beta-hydroxybutyrate (BHB), acetoacetate, and acetone), and / or cytokines (such as tumor necrosis factor (TNF) (e.g., TNF-α), interleukins (e.g., IL-6)). However, as noted above, the sensors may be placed in direct contact with the patient rather than in the blood circuit 112. Where they are present, the sensors are operatively connected to the computing device 12 via a wireless connection or a physical connection.

[0178] like Figure 5 As shown in schematic form in FIG, the blood circuit 112, including the inlet blood line 114 and the return blood line 116, can also include a thermal management system 500 for heating or cooling the blood within the blood circuit 112. The thermal management system 500 is operably connected to the computing device 12, 64. Such a thermal management system can be used to heat the blood returning to the patient, such as to about 37°C. Such a system can also be used to cool the returning blood as low as 20°C or to increase the temperature of the returning blood to 43°C, thereby warming the patient to a temperature as high as 38°C to 40°C. Furthermore, the dialysate circuit can also include a second thermal management system 501 to ensure that the blood has reached the desired temperature after passing through the blood filter 120.

[0179] Figures 3 and 4 Also shown is the dialysate fluid side or filtrate side of the system 110, which generally includes a dialysate fluid circuit 140 having a first dialysate fluid line segment 141 and a second dialysate fluid line segment 142. Each of these lines is connected to the filtration unit 120 on the side of the membrane opposite the segments 114, 116 of the blood circuit 112. Figures 3 and 4 In FIG. 1 , a respective fluid pump 144 , 146 is operatively associated with each of these pipe sections 141 and 142 .

[0180] The first tubing segment 141 is also connected to a dialysate fluid source (e.g., fluid bag 149), which may include electrolytes or other treatment compounds premixed therein. The second tubing segment 142 is connected to a waste collection device (e.g., a waste container such as bag 153). A pressure sensor 154 may also be disposed in the second dialysis fluid tubing segment 142.

[0181] Figures 3 and 4 A system is shown that is typically used as a base model for many dialysis procedures. Additional fluid lines, circuits, and components can be added (or deleted) to increase treatment options. In particular, in the present invention, additional lines can be provided for supplying (such as by adding to the blood circuit 112) ketones (e.g., hydroxybutyrate (BOHB), acetoacetate, and / or acetone) to the patient. Supplying ketones to the patient is important because it reduces the requirement for the patient to be in a state of meal-induced ketosis before treatment begins. In embodiments where a separate supply of ketones is not provided, the fluid source bag 149 can also include one or more ketones (such as sodium BOHB) in solution for diffusion through the membrane into the blood circuit.

[0182] One or more of the dialysate fluid source 149, waste container 153, and replacement fluid container 168 may be provided on a scale 400 in operable communication with the computing device 12. Any additional source of ketones may also be provided on such a scale 400. This allows the quality of the fluid in each container to be monitored during use to provide an accurate record of the amount of each fluid used or collected.

[0183] In addition, if Figures 3 and 4 As shown, the system 110 includes an extracorporeal blood control device 160 that provides a variety of treatment options and can be controlled and / or monitored via a control screen / display 161 (e.g., a control device or controller disposed in the system housing 193). Touch screen controls can be incorporated herein and / or other conventional knobs or buttons (not shown) can be used (e.g., a graphical user interface can be displayed via a touch screen as described herein). Additional and more detailed information about example devices 160 can be found in U.S. Patent Nos. 5,679,245; 5,762,805; 5,776,345; and 5,910,252, among others.

[0184] For illustrative purposes, a general description will be given of the Figures 3 and 41 and 2. A general dialysis treatment procedure is performed by the device described. First, blood is removed from the patient 118 via the access device 117 by, for example, a blood pump 124, and flows through the access line 114 to the filter 120. The filter 120 processes the blood according to one or more extracorporeal blood treatment profiles selected from a plurality of extracorporeal blood treatment profiles (e.g., selected and controlled by the screen interface 161 of the control device 160).

[0185] The treatment profile also involves reducing glutamine and glucose in the patient's blood. The concentration of glutamine is reduced to a value within the range of 0.1mM to 0.5mM, and preferably to a value within the range of 0.15mM to 0.3mM. The blood concentration of glucose is reduced to a concentration within the range of 1mM to 6mM, and preferably within the range of 2mM to 4mM. Since the glycolysis and glutaminolysis pathways are enhanced in many cancer cells, this treatment has a more significant and adverse effect on cancer cells than on healthy cells in the body. Although glycolysis is enhanced to meet the increased energy needs of cancer cells, glutaminolysis is also enhanced in many cancer cells and can provide biosynthetic precursors, but it also plays an important role in maintaining reactive oxygen species (ROS) in hemostasis. Forcing cancer cells into apoptosis by increasing ROS production in cancer cells is an important goal of radiotherapy and chemotherapy. Finally, glutamine can serve as an ATP source during periods of low glucose levels. In some embodiments, the treatment profile ensures that the patient's nutritional needs can be met by providing a suitable parenteral nutrition composition to the blood line or as a separate infusion directly into the patient's vascular system. In some embodiments, during treatment, the blood concentration of ketone bodies is maintained at a value within the range of 1 mM to 15 mM, and preferably within the range of 2 mM to 12 mM. Although patients are expected to produce their own ketone bodies (e.g., from ingested or infused lipids or stores of body fat) when exposed to such low blood glucose concentrations, the biological processes that produce sufficient amounts of ketone bodies may take time to begin. The introduction of ketone bodies during the treatment described herein ensures that even in the event that blood glutamine and blood glucose concentrations decrease at an abnormally high rate, the patient has a sufficient energy source for continued vital functions in a timely manner.

[0186] In certain treatments (particularly those that maintain elevated blood ketone concentrations), the patient is provided with a supply of one or more pharmacological agents to help lower blood glucose concentrations. Such glucose-lowering agents preferably include biguanides, α-glucosidase inhibitors, SGLT2 inhibitors, and dopamine agonists. Such agents can be supplied to the patient by the system by inclusion in the dialysate fluid source 149, an additional ketone source (if present), or as an additional infusion to be supplied to the blood circuit 112, or otherwise.

[0187] An example biguanide is metformin. Example α-glucosidase inhibitors include acarbose or miglitol. Example SGLT2 inhibitors include canagliflozin, dapagliflozin, and empagliflozin. Example dopamine agonists include bromocriptine.

[0188] In some preferred treatments, antiglycolytic agents may be provided to the patient to further inhibit glycolytic activity in tumor cells. Such agents may be provided to the patient by the system by inclusion in the dialysate fluid source 149, an additional ketone source (if present), or as an additional infusion to be supplied to the blood circuit 112, or otherwise.

[0189] Following processing, the system returns the processed or treated blood to the patient 118 via a return line 116 and a return device 119 , which are inserted into or otherwise connected to the vascular system of the patient 118 .

[0190] The blood flow path to and from the patient 118 forms a blood flow circuit 112, which includes an inlet device 117, an inlet line 114, a blood pump 124, a filter 120, and a return line 116 back to the patient and a return device 119. Pressure sensors can be used to sense various pressures in the system 110. For example, a pressure sensor 127 can be connected in the inlet line 114 and allow monitoring of the fluid pressure in the inlet line 114, and a second pressure sensor 128 can be connected in the blood circuit 112 between the first blood pump 124 and the blood inlet into the filter 120 and can be used to detect and monitor the pressure of the blood supplied to the inlet of the filter 120.

[0191] System 110 can also be included in the degassing chamber 125 in the return line, to provide the same operation as eddy current to push air out of the delivery path of blood. Replacement solution can be added in the degassing chamber on the top of blood after filtration, to prevent air / blood contact surface. Degassing chamber monitoring line 191 can use connection equipment (such as, return pressure port 129) to connect degassing chamber 125 to the internal pressure transducer in system housing 193. This enables return pressure monitoring to be achieved, and if necessary, is to remove air from degassing chamber and can be achieved. The return clamp 131 being connected in blood circuit 112 selectively allows or terminates blood to flow through blood circuit 112 (for example, whenever bubble detector 126 detects air in blood, return clamp 131 can be activated).

[0192] Additionally, pump 162 may be connected to anticoagulant container 164 to deliver anticoagulant to the blood in tubing segment 114 via anticoagulant line 165, and pump 166 may deliver replacement fluid from replacement fluid container or bag 168 via replacement fluid line 170. Secondary flow circuit 140 is also Figures 3 and 4 1 as it interacts with the filter 120. A secondary flow circuit 140 is connected to the secondary chamber of the filter 120. Substances removed from the blood outside the body are removed from the secondary chamber of the filter 120 via an outlet tube section 142 of the secondary flow circuit 140, and substances added to the blood outside the body move into the filter 120 via an inlet tube section 141 of the secondary flow circuit 140.

[0193] Secondary flow circuit 140 generally includes a fluid source such as bag 149, an inlet fluid line 141, a third pump 144, a secondary chamber of filter 120, a waste fluid line 142, a pressure sensor 154, a fourth pump 146, and a waste collection device such as container 153. Source fluid bag 149 can contain a sterile dialysate fluid that is generally isotonic with blood into which blood impurities diffuse through the semipermeable membrane of filtration unit 120. The fluid source bag can also include one or more ketones in solution for diffusion through the membrane into the blood circuit.

[0194] A pump 144 is connected in the inlet fluid line 141 for delivering dialysate fluid from a dialysate fluid source 149 to the inlet of the filter 120. A waste collection container 153 is provided to collect or contain material from the blood that is transferred across the semipermeable membrane in the filter 120 and / or to contain used dialysate fluid after it has passed through the filter 120. A fourth pump 146 is connected to the waste collection line 142 for moving used dialysate from the filter 120 to the waste collection container 153. A pressure sensor 154 may also be located in the waste collection line 142 for the purpose of monitoring the pressure in the secondary chamber of the filter 120.

[0195] The filtration unit 120, flow lines, and other components of the primary flow circuit 112 and secondary flow circuit 140 described herein (except, for example, a pump and possibly a few other objects) can be formed into an integral replaceable unit (e.g., an extracorporeal blood set). This integral replacement unit may be referred to herein as a "treatment device." An example of such a treatment device or integral replaceable unit is described in more detail in U.S. Patent No. 5,441,636, entitled "Integrated Blood Treatment Fluid Module" (see also U.S. Patent No. 5,679,245, entitled "Retention Device for Extracorporeal Treatment Apparatus").

[0196] Depending on the system configuration, treatment devices used in performing different treatments may be available. Figures 3 and 4As will be generally understood, the integrated tubing and filtration module (identified by reference numeral 172) includes the filter 120 and all of the tubing and associated components described above that can be connected to the device 160. For example, the filter and tubing can be held on a plastic support member 174, which can in turn be connected to the device 160 (e.g., connectable to the system housing 193 of the device 160). The treatment kit can also include sensors that monitor the concentration of glucose and / or glutamine; ketones (such as beta-hydroxybutyrate (BHB), acetoacetate, and acetone) and / or cytokines (such as tumor necrosis factor (TNF) (e.g., TNF-α), interleukins (e.g., IL-6)). Such sensors can be configured to monitor the concentration of the listed components in the blood circuit or can be configured to monitor the concentration of the listed components through direct contact with the patient's body.

[0197] When in the operative position connected to the apparatus 160, the flexible fluid conducting tubing lines to and from the filtration unit 120 are held in a loop in operative pump communication for operative contact with the peristaltic pumping components of the pumps 124, 144, 146, and 166 to cause fluid to flow through the primary (blood) circuit 112 and the secondary (dialysate fluid) circuit 140. The module 172, including the filter 120 and all tubing and associated flow components, may be disposable after use.

[0198] The peristaltic pump components of pumps 124, 144, 146, and 166 can be fixedly disposed on device 160 (eliminating the need for disposable tubing loop components) and can be reusable. Typically, electrical, mechanical, or electromechanical components are also fixedly disposed in or on device 160 (e.g., system housing 193 connectable to device 160). Examples of such components include display screen 161 (e.g., a touch screen), bubble detector 126, tubing clamp 131, and connection devices for coupling to pressure sensor devices implementing pressure sensors 127, 128, 129, 154. Connections can also be provided for any desired or preferred sensors.

[0199] As explained above, the entry device 117 and the return device 119 may comprise catheters. In some embodiments, the catheters may comprise occlusion catheters, such as balloon occlusion catheters. This arrangement allows for Figure 4 The illustrated approach delivers entry device 117 and return device 119 to the site of the tumor (e.g., via femoral artery or vein access). This allows for localized reduction of blood glucose concentrations around the tumor, thereby providing the potential to reduce blood glucose and / or glutamine concentrations to even lower levels.

[0200] Reference Example 1

[0201] The sensitivity of different human cancer cell lines to glucose, glutamine, and ketones in cell culture media was studied in the presence of concomitant depletion of selected nutrients, mimicking conditions obtained by cancer dialysis.

[0202] Study 1 was conducted on a selection of human cancer cell lines identified from renal cell carcinoma, colon cancer, and glioblastoma. First, the effects of growth in the presence of increasing concentrations of β-hydroxybutyrate on cell viability were investigated, along with limiting glucose and glutamine levels. The addition of citrate to the cell culture medium was also tested. Cells were cultured under these conditions for 3 days, after which cell viability was determined. In the first study, a major effect on cell viability was observed when glutamine was depleted from the culture medium.

[0203] Materials and methods

[0204] Cell culture conditions

[0205] The cell line set up from human colon cancer (HCT15, NCI-H508 and COLO205), renal cell carcinoma (769-P, 786-O and RCC4) and glioblastoma (LN-18, A-172 and U-118MG) is selected for analysis.Except RCC4 and HCT15 purchased from Sigma-Aldrich (Merck, Germany), all cell lines are all available from American model culture collection bank (ATCC, LGC standard, UK).In addition, comprise the primary human renal cell carcinoma (RCC) cell separated from patient's nephrectomy in the research.Follow the culture condition recommended by American model culture collection bank (ATCC), i.e., growing cell in the DMEM culture medium adding 1mM sodium pyruvate, this sodium pyruvate is also added into RPMI-1640 culture medium, to keep condition more similar.

[0206] According to the recommendation from ATCC, 769-P, RCC4, LN-18, A-172, U-118MG and primary RCC cells were cultured in DMEM high glucose medium, while 786-O HCT15, NCI-H508 and COLO205 were cultured in RPMI-1640 medium. 1% penicillin-streptomycin and 10% calf serum were added to both. Cells were expanded and aliquots were frozen according to standard procedures.

[0207] Determine the optimal seeding density for each cell line in a 96-well plate according to the "Protocol for Optimizing Cell Seeding Density to Ensure Logarithmic Growth." The protocol is as follows:

[0208] • Prepare single cell suspension and measure cell count / viability.

[0209] Dilute cells to approximately 160,000 cells / ml in complete medium. Add 200 μL of cells to the top row of a 96-well plate. Aliquot 100 μL of complete medium into all other wells. A small number of medium-only control wells are required on each plate to serve as blanks.

[0210] Repeatedly, using a 12-well pipette, dilute the cell preparation into two aliquots down the plate: 100 μL of medium per 100 μL of cells in the next row. Then, add 50 μL of complete medium to all wells. Cover the plate.

[0211] • Incubate the plate overnight at 37°C, 5% CO2.

[0212] • Add 50 μL of fresh culture medium to the wells to achieve a final volume of 200 μL and incubate at 37°C, 5% CO2 for 72 hours.

[0213] • Viability was measured using the CellTiter-Glo assay according to the manufacturer's protocol.

[0214] • Plot the number of cells against the logarithm of the luminescence intensity to find the cell concentration that achieves logarithmic growth.

[0215] The CellTiter-Glo Luminescent Cell Viability Assay (Promega) was used as a readout for viability.

[0216] Study 1

[0217] For each cell line, the cell inoculation of the optimal number determined above was in 96 well plates at day 0. Next day, after adding the nutrients outlined in table 3 and file " plate overview " (plate overview), cells were cleaned in PBS, and culture medium was replaced with DMEM (Fisher Scientific) or RPMI-1640 culture medium (Saveen Werner) that does not contain glucose or L-glutamine. Three holes were treated for each condition. After 3d cultivation in test conditioned medium, along with changing culture medium every day, cell viability was determined using CellTiter-Glo viability test. Repeat the experiment three times for every kind of cell line.

[0218] Table 1. Cancer cell lines and culture media

[0219] kidney 769-P DMEM high glucose 786-O RPMI-1640 RCC4 DMEM high glucose colon HCT15 RPMI-1640 NCI-H508 RPMI-1640 COLO205 RPMI-1640 brain CRL2610(LN-18) DMEM high glucose A-172 DMEM high glucose U-118MG DMEM high glucose

[0220] Table 2. Normal culture medium contents of selected nutrients

[0221]

[0222] Table 3. Test conditions for Study 1

[0223]

[0224] The matrix shows the different combinations of growth conditions used in Study 1. *The amount of glucose is expressed as a percentage of the concentration present in the standard culture medium for each cell line. Where indicated, 1 mM citrate was added to the culture medium. The indicated nutrients were added to DMEM (Fisher Scientific) or RPMI-1640 medium (SaveenWerner) without glucose or L-glutamine.

[0225] Table 4 Product order information

[0226]

[0227]

[0228] The results of Study 1 are presented in the graph shown in Figure 6. A clear correlation was shown between reducing the presence of glutamine and cell culture medium and reducing cell proliferation.

[0229] Study 2

[0230] As described above, in Study 1, ATCC-recommended culture conditions were followed. However, pyruvate is a potential energy source that could affect the results. Therefore, in Study 2, the same culture conditions as in Study 1 were tested in two cell lines, A172 (glioblastoma) and RCC4 (renal cell carcinoma), in DMEM medium without the addition of sodium pyruvate. The results are presented in Figure 7. As in Study 1, a clear correlation was shown between reducing the presence of glutamine and the cell culture medium and reducing cell proliferation. However, in the absence of pyruvate in the cell culture medium, the results were much more significant. It also appears that increasing the concentration of BOHB ketone also inhibited cell proliferation.

[0231] All patents, patent documents, and references cited herein are incorporated in their entireties, as if each were individually incorporated. The present disclosure has been provided with reference to illustrative embodiments, and the present disclosure is not meant to be interpreted in a limiting sense. As previously described, those skilled in the art will recognize that various other illustrative applications can utilize the beneficial properties of the apparatus and methods described herein using the techniques described herein. Various modifications of the illustrative embodiments and additional embodiments of the present disclosure will be apparent upon reference to this specification.

[0232] Example 2

[0233] Study Design

[0234] Growth medium

[0235] In order to study the effect of a nutrient-restricted ketogenic environment on the growth of cancer cells in vitro, three different cell culture media were prepared. Culture medium A is a complete RPMI1640 culture medium in which cell lines are routinely cultured. Culture medium B is used as an approximation of the conditions found in normal human serum. The levels of glucose, glutamine, serine, glycine, and arginine are adjusted to match the normal physiological levels found in human serum. These nutrients are selected based on their reported use as energy sources and their effects on the metabolic state of cancer cells. Culture medium C is used to simulate cancer dialysis conditions that limit ketogenic nutrients. Here, the levels of selected nutrients are reduced to half of the physiological levels in culture medium B, and ketone bodies BOHB are added.

[0236] The composition of each culture medium is described in the manner of Materials and Methods listed in Tables 5 to 6.

[0237] oxygen levels

[0238] Human cancer cell lines are routinely defined and cultured at atmospheric oxygen levels (21% O2). However, physiological oxygen levels in tissues are quite low and vary from 3% to 13% [Ward, Biochim Biophys Acta 2008;1777:1-14]. Within the tumor microenvironment, the rapid growth rate of cancer cells combined with the often malformed and defective vasculature often results in hypoxic regions with oxygen levels ranging from 0 to 5%. Given the impact of oxygen levels on energy metabolism [Xie et al., J Biol Chem 2017;292:16825-16832], and to further mimic physiological conditions in vivo, the growth of cancer cell lines in media A, B, and C was studied in both 21% O2 and a more physiological 5% O2 environment.

[0239] ketone

[0240] Acetoacetate ketone (acac), BOHB, and acetone are produced by the liver during fasting or starvation. BOHB is the major ketone body in mammals, with acac constituting approximately 20%. Most published in vitro studies investigating the effects of ketones on cancer cells have focused primarily on BOHB; however, some studies have shown that the addition of acac has different effects compared to BOHB [Vallejo et al., J Neurooncol 2020;147:317-326]. To further simulate the in vivo ketogenic situation in which both ketones are present, and to investigate the possible differential effects of BOHB and acac, acac was also included in the study.

[0241] Materials and methods

[0242] cell lines

[0243] All cell lines were purchased from ATCC (ATCC, LGC standard) except RCC4 from Sigma-Aldrich (Merck). Primary human renal cell carcinoma cells were isolated from nephrectomies performed at Sahlgrenska University Hospital in Gothenburg, Sweden, after informed consent from the patients and approval by the regional ethics committee. The optimal seeding density for each cell line was determined in 96-well plates cultured for 3 days in standard cell culture medium.

[0244] Culture conditions and additives

[0245] Cells were maintained in RPMI-1640 medium (31870-025GIBCO) supplemented with 10% serum, 200 mM L-glutamine, and 1% penicillin-streptomycin (PEST) in a humidified chamber at 37°C and 5% CO2. For hypoxic conditions (5% O2), cells were maintained in a Galaxy 14S CO2 incubator (Eppendorf) with the O2 level adjusted to 5% using N2.

[0246] Culture media A, B, and C were prepared as follows.

[0247] Culture medium A: RPMI1640 (31870-025, GIBCO) supplemented with 1% PEST, 200 mM L-glutamine, and 10% dialyzed serum. Dialyzed serum is used to reduce the amount of small molecules such as amino acids.

[0248] Substratum B and C are prepared by the culture medium powder (R9010-01, USBiologicalLife Sciences) of no L-glutamine, glucose and amino acid whose RPMI1640 is improved.For 1L substratum, 7.4g powder is dissolved in 900mL sterilized water when not heating, and 2g sodium bicarbonate is added.The amino acid listed in table 5 is added into and the concentration (table 5) identical in complete RPMI1640 substratum.After all additions, by means of by 0.22um membrane filtration by substratum sterilization, and be divided into two bottles.

[0249] In Medium B, to mimic physiological conditions, the levels of glutamine, serine, glycine, arginine, and glucose were set to the median of the levels measured in human serum based on data from Mayo Clinic Laboratories (https: / / www.mayocliniclabs.com / test-catalog / clinical+and+interpretive / 9265).

[0250] To model cancer dialysis conditions in Medium C, the levels of these nutrients were reduced to 50% of physiological levels. For Medium A, 1% PEST and 10% dialyzed serum were added to Mediums B and C. The concentrations of selected nutrients in Mediums A to C are summarized in Table 6. Sodium pyruvate, a common additive in cell culture media, was not present in any of the media used.

[0251] Table 5. Amino acid concentrations in RPMI1640

[0252]

[0253] Table 6. Nutrient composition of culture media A, B and C

[0254]

[0255]

[0256] Amino acids and other additives were purchased from Sigma Aldrich.

[0257] After all nutrients were added, the pH was measured. The pH values ​​were as follows: complete RPMI 1640 with 10% non-dialyzed FBS, 1% PEST, and 200 mM L-glutamine, pH 7.78; Medium A, pH 7.56; Medium B, pH 7.62, and Medium C, pH 7.57.

[0258] Stock solutions of DL-β-hydroxybutyrate sodium salt (H6501, Sigma Aldrich) and lithium acetoacetate (A8509, Sigma Aldrich) were prepared in water, sterile filtered, aliquoted and stored at −20° C. Lithium chloride (L7026, Sigma Aldrich) was used as a control for the addition of lithium to Li-Acac.

[0259] Viability assay

[0260] The CellTiter-Glo Luminescent Cell Viability Assay (Promega) was used as a readout of cell number according to the manufacturer's instructions. Figures 12 to 13 In the experiment shown, two plates were inoculated and processed. One set of plates was used to collect culture medium for lactate measurement (see below) and CellTiterGlo assay. At the end of the experiment, another set of plates was frozen at -80°C. The frozen plates were intended for CyQuant cell proliferation assays (Thermo Fisher), which measure the amount of DNA in each well. Analyzing cell amounts by both CellTiterGlo and CyQuant assays will ensure that the effects of culture conditions on viability or growth rate will not be masked by simultaneous changes in ATP levels per cell.

[0261] Collecting culture medium for lactate measurement

[0262] exist Figures 12 to 13 In the experiment shown, the cell culture medium was collected on day 3, transferred to a new 96-well plate and frozen at -80°C. This culture medium can be used to analyze the amount of secreted lactate as a measure of metabolic state. Several kits for lactate measurement are available, for example, the Lactate-Glo assay (J5021, Promega) is designed for use in the presence of serum.

[0263] result

[0264] Example 2 is designed to answer the following questions:

[0265] - Is the growth of selected cancer cell lines affected by simulated cancer dialysis conditions in Medium C under normoxia or hypoxia?

[0266] Growth in media A, B, and C

[0267] As a first step, Medium A to Medium C were prepared as described in Materials and Methods, and the cancer cell lines were tested for their ability to grow in these media. Growth curves of the selected cell lines were determined over time in each medium. Cell counts were analyzed after 1, 2, and 3 days of culture in Medium A to Medium C under normoxia (21% O2).

[0268] like Figure 10 As shown, reducing selected nutrients to less physiological levels as in Medium B significantly reduced the growth rate of the A549 lung cancer cell line and the RCC4 renal cancer cell line compared to Medium A. Medium C further reduced the growth rate compared to Medium A.

[0269] Growth of cancer cells in medium A to medium C under normoxia and hypoxia

[0270] exist Figure 11 Figure 2 shows the cell mass of A549 lung cancer cell line and RCC4 renal cancer cell line, as well as primary renal carcinoma (RCC) cells, after 3 days of culture in medium A to medium C under normoxia and hypoxia. Furthermore, the growth rate in mediums B and C under normoxia was reduced compared to medium A. The same pattern was observed in cells cultured under 5% O2. Furthermore, in RCC4, there was no significant additional effect in medium C with low nutrient levels and the addition of BOHB compared to the conditions in medium B.

[0271] For A549, a small but significant decrease in growth rate was found between media B and C, but only under normoxia.

[0272] This study included primary renal carcinoma cells from three patients. Similar to established cell lines, these cells showed reduced growth rates in Medium B and C compared to Medium A.

[0273] In conclusion, changing the oxygen pressure from 21% O2 (normoxia) to 5% O2 (hypoxia) had a very limited effect on the growth rate of these cells.

[0274] Next, it was decided to also include Acac in the study and analyze the viability of cells cultured in the presence of BOHB and Acac, alone or in combination, in Medium A to Medium C. The experiments were performed under 21% O2 and 5% O2. The experiments were performed using the glioma cell lines A172 and U118MG.

[0275] As a chiral molecule, BOHB exists as two enantiomers, D-BOHB and L-BOHB. D-BOHB is normally produced and metabolized in humans. The BOHB salt used in this study contained a 50:50 mixture of D-BOHB and L-BOHB. To ensure the presence of the highly active D-form, the total concentration of added BOHB was increased to 16 mM, resulting in a level of 8 mM D-BOHB. To maintain a constant total concentration of active ketones, 8 mM Acac was used, and for combinations of both ketones, the levels were adjusted to 4 mM Acac and 8 mM BOHB (containing 4 mM D-BOHB).

[0276] Acac is available for in vitro use in the form of a lithium salt. Because lithium itself can affect the viability of cancer cells [Cohen-Harazi et al., Anticancer Res 2020;40:3831-3837], 8 mM LiCl was used as a control for the 8 mM Acac data points, and 4 mM LiCl was used as a control for the 4 mM Acac / 8 mM BOHB data.

[0277] At the end of the experiment, the culture medium from each well was collected and frozen to enable later determination of lactate levels as a measure of metabolic state. In addition, a duplicate experiment was performed in which one set of plates was frozen for later quantification of cell number using the CyQuant proliferation assay, and the cell amount in the other set was analyzed by the CellTiterGlo viability assay.

[0278] Effects of BOHB and Acac added alone or in combination

[0279] like Figures 12 to 13 As shown, initial experiments gave promising data on the effects of high ketone concentrations on the growth of A172 and U118MG glioma cell lines under nutrient-reduced conditions.

[0280] In medium A, the addition of 16 mM BOHB alone had no growth inhibitory effect on A172 or U118MG cells under normoxia and hypoxia, and 8 mM Acac did not further reduce cell number compared with the 8 mM LiCl control.

[0281] However, the addition of 4 mM Acac in combination with 8 mM BOHB significantly reduced the number of A172 cells in normoxic Medium A compared to the 4 mM LiCl control.

[0282] The interpretation of the results from medium B is confounded by technical errors in normoxic control samples. However, in hypoxic A172 cells, BOHB significantly reduced cell numbers to approximately 70% of the amount in medium B without BOHB. A similar reduction was observed in hypoxic U118MG cells.

[0283] Furthermore, in medium B, 8 mM Acac significantly reduced cell number compared to the 8 mM LiCl control in both cell lines, but only at 21% O2.

[0284] In medium C, the addition of 8 mM Acac alone did not significantly reduce cell mass compared to the 8 mM LiCl control. However, in both cell lines and at 21% O2 and 5% O2, the addition of 16 mM BOHB significantly reduced cell number to approximately 30% compared to medium C without BOHB.

[0285] Furthermore, in both cell lines and at both oxygen levels, the combination of BOHB and Acac resulted in significantly fewer cells in Medium C compared to the 4 mM LiCl control.

[0286] These results show that high levels of BOHB or Acac alone cannot inhibit the growth of glioma cells in a nutrient-rich environment (such as culture medium A). In addition, in culture medium B with more physiological nutrient levels, when ketones were added, only small differences were observed. The greatest impact was found in culture medium C, where, under both hypoxia and normoxia, the combination of 16mM BOHB alone and 8mM BOHB with 4mM Acac significantly reduced cell number compared to their respective controls. When 8mM Acac was added alone, this was not observed.

[0287] However, repetition of these experiments, focusing on media B and C under normoxic conditions, gave inconsistent results. Figures 14 to 16Shown are the results for the combination of glioma cell lines A172 and U118MG and renal cancer cell line RCC4 from Experiments 2 to 6. When BOHB or Acac was added alone, a trend toward decreased growth was observed in Medium C, particularly in the A172 cell line.

[0288] The results from the first and second parts of Example 2 indicate that cancer cell lines grow slower in a nutrient-limited environment. The cell lines tested appeared viable in media B and C, albeit with reduced proliferation rates. Optical examination of the cells on day 3 did not reveal any floating cells, which could be a sign of dying cells.

[0289] Data from the third part of the study showed increased sensitivity of glioma cell lines to high levels of BOHB or a combination of BOHB and ACAC in the nutrient-limited medium C. However, no such sensitivity was found for the renal cancer cell line RCC4.

Claims

1. An extracorporeal blood processing system (50) for treating a subject suffering from cancer, the system comprising: Extracorporeal blood circuit (52a, 52b); a dialysate fluid circuit (54a, 54b); The extracorporeal blood circuit (52a, 52b) and the dialysate fluid circuit (54a, 54b) are separated by a membrane (59) of a filtration unit (58); at least one blood pump (60) for controlling the flow of blood through the extracorporeal blood circuit (52a, 52b); at least one dialysate fluid pump (62, 68) for controlling the flow of dialysate fluid through the dialysate fluid circuit (54a, 54b); a system computing unit (64) operatively connected to the blood pump (60) and the dialysate fluid pumps (62, 68), the system computing unit having a user interface comprising an input device and a display device; wherein, The system calculation unit (64) is adapted to receive a desired blood concentration value GLN of nitroglycerin in the range of 0.1 mM and 0.5 mM. b ; The system calculation unit (64) is adapted to receive a desired blood concentration value GLUCOSE of glucose in the range of 2 mM to 4 mM b ; The system calculation unit (64) is adapted to receive a desired blood concentration value KETONE of ketone bodies in the range of 1 mM to 15 mM. b ; The system computing unit (64) is adapted to receive a concentration value GLN p , the concentration value GLN p represents the concentration of glutamine or a pharmaceutically acceptable glutamine-containing compound in fresh dialysate fluid; The system computing unit (64) is adapted to receive the concentration value GLUCOSE p , the concentration value of GLUCOSE p represents the concentration of glucose in fresh dialysate fluid; The system calculation unit (64) is adapted to receive the actual glutamine concentration value GLN in the blood of the subject being treated. a , and adapted to receive the actual glucose concentration value GLUCOSE in the blood of the subject being treated a , and is suitable for receiving the actual concentration value of ketone bodies KETONE a ; The system computing unit (64) is adapted to control the blood pump (60) and the dialysate fluid pumps (62, 68) such that the actual blood concentration value GLN of glutamine is a Driven close to or below GLN b , and the actual blood concentration of glucose GLUCOSE a Driven close to or below GLUCOSE b ; and, In case the system (50) comprises one or more of the infusion lines (66, 80, 81, 82), and in case one of the infusion lines (66, 80, 81, 82) is adapted to infuse an infusion liquid into an extracorporeal blood line (52b) or directly into the vascular system of the subject to be treated, the system computing unit (64) is adapted to control the infusion pump of the infusion line such that the actual blood concentration value KETONE a Driven close to KETONE b ;or In case the system (50) does not include the infusion line (66, 80, 81, 82), the system computing unit (64) is adapted to convert the KETONE a with KETONE b Compare and compare in KETONE a <KETONE b In the case of the above, it is suitable to display a message on the display device to inform the subject being treated that he should consume more ketone bodies or medium-chain triglycerides.

2. The extracorporeal blood treatment system (50) according to claim 1, wherein: The system computing unit (64) is adapted to receive the concentration value KETONE p,, , the concentration value KETONE p,, Represents the concentration of ketone bodies in fresh dialysate fluid.

3. The extracorporeal blood treatment system (50) according to claim 1, wherein: The system (50) comprises one or more of the infusion lines (60, 80, 81, 82), each of the infusion lines being connected to the extracorporeal blood circuit (52a, 52b) or adapted to be directly connected to the vascular system of the subject to be treated, each of the infusion lines comprising an infusion pump; the system computing unit being operatively connected to the one or more infusion pumps of the one or more infusion lines (66, 80, 81, 82); and The system computing unit (64) is adapted to receive a concentration value KETONE via one of the one or more infusion lines (66, 80, 81, 82) i , the concentration value KETONE i Indicates the concentration of ketone bodies in the infusion fluid to be infused into the extracorporeal blood line (52b) or directly into the vascular system of the subject to be treated.

4. The extracorporeal blood treatment system (50) according to claim 3, wherein: The system computing unit (64) is adapted to monitor GLN a , and in GLN a Lower than GLN b In the case of GLN, it is suitable to start the infusion of the composition comprising glutamine or a pharmaceutically acceptable compound comprising glutamine by activating the associated infusion pump of one of the one or more infusion lines (66, 80, 81, 82), and to maintain the infusion until GLN a Equal to GLN b .

5. The extracorporeal blood treatment system (50) according to claim 3, wherein: The system computing unit (64) is adapted to monitor GLUCOSE a , and in GLUCOSE a Lower than GLUCOSE b In the case of a condition in which the patient is taken into consideration, the patient is adapted to start infusing the composition comprising glucose by activating the associated infusion pump in one of the one or more infusion lines (66, 80, 81, 82), and to maintain the infusion until the patient is taken into consideration. a Equal to GLUCOSE b .

6. The extracorporeal blood treatment system (50) according to claim 4, wherein: The system computing unit (64) is adapted to monitor GLUCOSE a , and in GLUCOSE a Lower than GLUCOSE b In the case of a condition in which the patient is taken into consideration, the patient is adapted to start infusing the composition comprising glucose by activating the associated infusion pump in one of the one or more infusion lines (66, 80, 81, 82), and to maintain the infusion until the patient is taken into consideration. a Equal to GLUCOSE b .

7. The extracorporeal blood treatment system (50) according to any preceding claim, wherein The membrane has a molecular weight cut-off (MWCO) of less than 60 kDa.

8. The extracorporeal blood treatment system (50) according to claim 7, wherein: The membrane has a MWCO of less than 50 kDa or less than 40 kDa.

9. The extracorporeal blood treatment system (50) according to claim 8, wherein: The membrane has a MWCO of less than 30 kDa, or less than 10 kDa, or less than 5 kDa, or less than 2 kDa.

10. The extracorporeal blood treatment system (50) according to any one of claims 1 to 6 and 8 to 9, wherein: The extracorporeal blood circuit comprises a thermal management system for heating or cooling the blood in the blood line during use.

11. The extracorporeal blood treatment system (50) according to claim 10, wherein: The thermal management system is controllable to regulate the temperature of the blood in the extracorporeal blood circuit to a temperature between 20°C and 43°C.

12. The extracorporeal blood treatment system (50) according to any one of claims 1-6, 8-9 and 11, further comprising one or more sensors (S, 90, 91, 92, 93) for detecting an analyte selected from the group consisting of glucose, glutamine and ketone bodies, said sensors (S, 90, 91, 92, 93) being positioned in the effluent portion (54a) of the dialysate fluid circuit, said sensors being in communication with the system computing unit (64) and providing an output indicative of the concentration of said analyte in the blood or in the spent dialysate fluid; wherein The system computing unit (64) is configured to determine a representative blood concentration of the analyte from the output of the sensors (S, 90, 91, 92, 93) to monitor GLN a 、GLUCOSE a and KETONE a At least one of .

13. The extracorporeal blood treatment system (50) according to any one of claims 1-6, 8-9 and 11, wherein: The ketone bodies include acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of β-hydroxybutyrate.

14. The extracorporeal blood treatment system (50) according to claim 1, wherein: A dialysis fluid for use in the extracorporeal blood treatment system (50) comprises ketone bodies.

15. The extracorporeal blood treatment system (50) according to claim 14, wherein The dialysis fluid further comprises at least one of the following: a) glutamine or a compound comprising glutamine; and b) Glucose.

16. The extracorporeal blood treatment system (50) according to claim 15, wherein a) the concentration of glutamine or a compound comprising glutamine is in the range of 0 mM to 0.5 mM; b) the concentration of glucose is in the range of 0 mM to 6 mM; and c) The concentration of ketone bodies is in the range of 1 mM to 15 mM.

17. The extracorporeal blood treatment system (50) according to claim 16, wherein: a) the concentration of glutamine or a compound comprising glutamine is in the amount of 0.05 mM to 0.3 mM; b) the concentration of glucose is in the range of 0.5 mM to 4 mM; and c) The concentration of ketone bodies is in the range of 2 mM to 12 mM.

18. The extracorporeal blood treatment system (50) according to any one of claims 14 to 17, wherein: The ketone bodies include acetoacetate, β-hydroxybutyrate or pharmaceutically acceptable derivatives, esters and salts of β-hydroxybutyrate.

19. A treatment device for use in an extracorporeal blood treatment system (50) according to any one of claims 1 to 18, the device comprising: A filter unit (58) having a membrane (59) separating an integrated blood line (52a, 52b) and an integrated dialysate fluid line (54a, 54b), wherein the blood line (52a, 52b) and / or the dialysate fluid line (54a, 54b) comprises a sensor (S, 90, 91, 92, 93) for monitoring GLN a 、GLUCOSE a and KETONE a At least one of .

20. A system computing unit (64) adapted to control an extracorporeal blood treatment system (50) for treating a subject suffering from cancer, the system computing unit comprising: a plurality of output devices adapted to be operatively connected to at least one blood pump (60), at least one dialysate fluid pump (62, 68); a user interface, including an input device and a display device; as well as Memory devices and computing devices; The system computing unit (64) is adapted to receive a desired blood concentration value GLN of glutamine b ; The system calculation unit (64) is adapted to receive a desired blood concentration value of glucose GLUCOSE b ; The system computing unit (64) is adapted to receive a desired blood concentration value of ketone bodies KETONE b ; The system computing unit (64) is adapted to receive the dialysate concentration value GLN of glutamine p ; The system computing unit (64) is adapted to receive a dialysate concentration value of glucose GLUCOSE p ; The system calculation unit (64) is adapted to receive the actual blood concentration value GLN of glutamine a ; The system calculation unit (64) is adapted to receive the actual blood concentration value of glucose GLUCOSE a ; The system calculation unit (64) is adapted to receive the actual blood concentration of ketone bodies KETONE a ; The system computing unit (64) is adapted to control the blood pump (60) and the dialysate fluid pumps (62, 68) such that the actual blood concentration value GLN of glutamine is a Driven close to or below GLN b , and the actual blood concentration value of glucose GLUCOSE a Driven close to or below GLUCOSE b ; and, In case that the infusion pump is operatively connected to the system computing unit (64), the system computing unit (64) is adapted to control the infusion pump so that the actual blood concentration value KETONE a Driven close to KETONE b ; and, In the absence of an infusion pump operatively connected to the system computing unit (64), the system computing unit (64) is adapted to convert the KETONE a with KETONE b Compare and compare in KETONE a <KETONE b In the case of the above, it is suitable to display a message on the display device to inform the subject being treated that he should consume more ketone bodies or medium-chain triglycerides.

21. The system computing unit (64) of claim 20, wherein: The plurality of output devices are adapted to be operably connected to one or more infusion pumps for controlling flow in each of the one or more infusion lines (66, 80, 81, 82).

22. The system computing unit (64) of claim 20, wherein: The system computing unit (64) is adapted to receive the concentration value KETONE p,, , the concentration value KETONE p,, Represents the concentration of ketone bodies in fresh dialysate fluid.

23. The system computing unit (64) of claim 20, wherein: The system computing unit (64) is adapted to receive a ketone body infusion fluid concentration value KETONE i。 24. The system computing unit (64) according to any one of claims 20 to 23, wherein: The ketone bodies include β-hydroxybutyrate and / or acetoacetate.

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