Patent Application
20240044763
The present invention relates to a method of simulating the dynamic interplay between gastrointestinal release, dissolution, and/or digestion and subsequent intestinal absorption of substances and/or compounds, in particular pharmaceutical substances and/or compounds. The present invention further relates to a gastrointestinal tract simulation system, a compartment for such a system, and a method of operating such a system. The gastrointestinal tract simulation system can be a simulator for any monogastric animal (including humans).
The conventional SHIMS is a dynamic model of the human gut comprising compartments respectively simulating the stomach, small intestine and ascending, transverse and descending colon. The stomach and small intestine compartments mimic the enzymatic and physicochemical environment by controlling pH and residence time and the dosing of a proper nutritional medium, enzymes and bile salts.
By controlling the pH, redox potential and residence times, the different colon compartments each harbor a microbial community that corresponds to that of the in vivo situation in terms of metabolic activity and community composition. As such, this system is a validated in vitro tool to perform pre-clinical research about the bidirectional interaction between orally ingested substances and the colonic microbiota activity and composition. However, this model was not designed and optimized to generate in vivo predictive pre-clinical data about the gastrointestinal disintegration, dissolution, and/or digestion and subsequent absorption of substances and more particular pharmaceutical substances and/or compounds.
A large body of new chemical entities suffer from very low aqueous solubility thereby resulting in a very low systemic exposure of the drug upon ingestion. Since only the dissolved fraction of a drug can be absorbed, pharmaceutical companies try to overcome this bioavailability-limiting factors by using several types of formulation strategies such as, among others, pH adjusting excipients, amorphous solid dispersions, surfactants, and cyclodextrins and/or dosing of the compound under fasted or fed state conditions (food effects) which have the ultimate goal to increase the amount dissolved drug in the small intestine thereby potentially increasing the percentage of the administered dose available for small intestinal absorption. In order to accelerate drug development pharmaceutical companies are more and more relying on pre-clinical data obtained through in vitro systems that are used to study the dissolution of pharmaceutical compounds. Static dissolution systems (no dynamics in concentration of tested compound in function of time, absence of dynamic pH conditions in stomach, absence of dynamic concentrations of solubilizing substances in the small intestine) can be used to obtain preliminary pre-clinical data about factors affecting the solubility of a pharmaceutical compound. Considering the highly dynamic nature of the gastrointestinal tract in terms of stomach emptying patterns (fed versus fasted state), fed state stomach pH conditions in function of time, and intercoupled dynamic concentrations of enzymes, bile salts, phospholipids in the small intestine in function of time, the use of simulatory systems, incorporating the dynamic interplay of all these environmental parameters could result in a more in-depth insights into the dissolution of pharmaceutical compounds along the gastrointestinal tract thereby potentially resolving research questions that could not be addressed using static systems. However, increasing the solute intraluminal small intestinal concentration of substances through specific formulation strategies or due to micellar solubilization under fed state conditions does not always result in increased absorption of the substance, a phenomenon which is known as the solubility-permeability interplay. Increasing the solubility of pharmaceutical compounds under fed state conditions or by using cyclodextrins or surfactants mostly rely on the entrapment and solubilization of the drug in bile salt micelles, surfactant micelles and cyclodextrin complexes thereby increasing the apparent solubility (total amount of detectable compound in solution) of the drug. However, to become available for absorption the drug needs to get out of these solubilizing drug structures and become molecularly dissolved. Depending on the formulation strategy and drug compound used the equilibrium between the solubilized (micelles and complexes) state and the molecularly dissolved (free compound) state can be favored towards the direction of the solubilized state, hence the increase in solute concentration will not be translated into increased absorption. Differentiating between both solute states of a drug can be performed using permeation barriers which can distinguish between drug complexes/micelles (solubilized state) and freely molecularly dissolved drugs. The use of static dissolution systems coupled with offline permeation devices could be used as a pre-clinical tool to study dissolution and permeation of a drug. However, due to the inherent dynamic nature of the dissolution of compounds in the gastrointestinal tract it is important to investigate the kinetics of the interrelated dissolution and permeation processes of a compound. Conditions in which the luminal concentration of the compound in continuously changing and the luminal compartment of the small intestine is directly coupled to the absorptive compartment. These dynamic interconnected dissolution and permeation processes are not simulated when using offline permeation tools or static coupled dissolution/permeation in vitro models. Next to this, these in vitro tools suffer from several drawbacks. In these systems the permeation of pharmaceutical compounds is tested by adding the compound to the donor side (simulating the intestinal lumen) of the device which is in contact with an acceptor solution (simulating the blood stream). In between both compartments a permeation barrier is present which only allows the molecularly dissolved fraction of the compound to permeate from the donor compartment to the acceptor compartment. Depending on the system, multiple factors can result in the absence of (sufficient) permeation in the device and the generation of biased data. Factors include:
Hence, an in vitro model that can accurately simulate the physiological conditions in the stomach and small intestine (pH and biorelevant media), and the dynamics of the gastrointestinal tract (gastric secretion, gastric emptying, dynamic stomach pH, duodenal secretion, small intestinal absorption, and small intestinal emptying) and that has an online permeation device is required to accurately simulate the dynamic gastrointestinal dissolution of a drug and its associated interconnected intestinal absorption (permeation). More specifically the online permeation compartment of the in vitro system should be optimized to overcome the drawbacks associated with the existing permeation systems (see above) and therefore should be designed in such a way that:
As such biorelevant dissolution of the compound is directly coupled to a permeation barrier that is designed in such a way that sufficient (detectable) permeation of a pharmaceutical compound during physiologically relevant time periods (biorelevant transit of dosage form) can be generated.
An object of the invention is to provide an improved method of simulating the (dynamically interconnected) gastrointestinal dissolution and intestinal permeation of substances, preferably pharmaceutical substances. Throughout the text, unless explicitly mentioned differently, the term “substance” and “compound” are preferably used interchangeably, as is the case for the term “absorption” and “permeation”. Another object of the invention is to provide an improved method of simulating the gastrointestinal dissolution and intestinal permeation of substances using a gastrointestinal tract simulation system.
This object is achieved by a method of simulating the gastrointestinal dissolution and intestinal permeation of a (pharmaceutical) substance according to independent claim 1.
In a first aspect, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal permeation of substances. For example, the substance may be pharmaceutical compounds, food components, chemically synthesized compounds, biological compounds, polysaccharides, proteins, peptides, amino acids, or lipids. The method comprises providing a dynamic gastrointestinal tract simulation system comprising at least two consecutive compartments of which a first compartment simulates (at least part of) the stomach and a second compartment simulates (at least part) of the small intestine, preferably the duodenum. To simulate the (at least part of) the stomach, the first compartment is at least partially, e.g. mainly, filled with simulated gastric fluids, i.e. a solution that simulates the composition and pH of gastric juice, and to simulate the (at least part of) the intestine, the second compartment is at least partially, e.g. mainly, filled with simulated intestinal fluids, also known as artificial intestinal fluid or synthetic intestinal fluid, i.e. a solution that simulates the components and pH of intestinal fluid.
In the second compartment, preferably in each compartment, the pH of the simulated fluid can be varied over time through active control via pH monitoring and pH setpoint adjustments using acid and base additions. Advantageously, the above method allows for independent pH control of the second compartment.
The gastrointestinal tract simulation system further comprises a fluid transfer system for transferring fluids into and from said at least two compartments. In particular, the fluid transfer system is configured to unidirectionally transfer fluids from the first compartment to the second compartment, and mutates mutandis from any one of the at least two compartments to the subsequent compartment.
The second compartment comprises an outer vessel and an inner vessel mounted inside the outer vessel, said inner vessel having a wall comprising a grid structure, preferably cylindrical, and a semi-permeable membrane, preferably a dialysis membrane, i.e. a semi-permeable film containing various sized pores, mounted around said grid structure. In contrast to ultrafiltration, in which drug-containing eluate flows across the surface of an ultrafiltration membrane of the membrane filter under pressure, the average flow rate of fluid transferred into the inner vessel is about equal to the average flow rate of fluid transferred out of the inner vessel so that no pressure builds up.
The term “semi-permeable membrane” as used herein is intended to designate a comparatively thin physical structure of such a character that certain dissolved compounds or substances with a molecular size below a certain molecular size cut off, depending on the pore size of the physical structure, can pass through it while others such as, for example, micellar and other colloidal drug structures and solid drug, having a molecular size above the specific molecular size cut off, cannot pass through it. Because of this meaning of the word semi-permeable membrane it is considered that any semi-permeable membrane can be defined as or considered as a form of a barrier which will allow passive diffusion of substances which have a size below the molecular size cut off of the membrane while substances with a greater size will be retained by the physical structure thereby preventing their passive diffusion.
The word “porous” indicates pore of any size or size range capable of retaining substances primarily because of the sizes of such substances.
The method further comprises the step of introducing the substance in simulated gastric fluids present in the first compartment, in a preferred embodiment, the method further comprises testing the concentration of total and dissolved substance in the first compartment through sampling of its content, transferring said gastric fluids from the first compartment into said inner vessel, testing the concentration of total and dissolved substance in said inner vessel through sampling of its content, and testing the flow of dissolved substance from said inner vessel by permeation through the semi-permeable membrane into the outer vessel through sampling of its content.
Advantageously, the above method allows to study the gastrointestinal dissolution and intestinal permeation of substances, in particular in oral dosage forms, in a dynamic dissolution model with a coupled and interconnected permeation setting. The above method allows to simulate the dissolution behavior of a substance (and optionally its specific formulation), the total and solute concentration of the substance in the stomach and small intestine, absorption of molecular dissolved substance over the intestinal wall, and the resulting concentration of the substance in the blood stream.
Contrary to permeation simulation systems that can only be used for offline assessment, i.e. under steady-state conditions, using a dynamic gastrointestinal tract simulation systems with coupled online permeation, allows to remove solute substance from the inner vessel, i.e. the small intestinal dissolution vessel, thereby reducing the solute substance concentration in the small intestine as occurring in vivo.
Contrary to pressure-driven processes, using a concentration-driven process, allows to more accurately simulate the permeation process in the small intestine as occurring in vivo.
Contrary to permeation systems an sich, using a system comprising at least two consecutive compartments with said second compartment having a semi-permeable membrane mounted around said cylindrical grid structure, allows to more accurately simulate the gastrointestinal tract, i.e. at least the stomach and the small intestine, by embedding the semi-permeable membrane mounted around said cylindrical grid structure in a dynamic in vitro gastrointestinal tract simulation system.
According to embodiments, the present invention relates to the method as described above, wherein the gastrointestinal tract simulation system is fully dynamic, i.e. a system that, during use, is continuously dynamic, meaning that liquid is approximately continuously pumped into the system and out of the system thereby simulating the continuous gastric secretion, gastric emptying, duodenal secretion, and/or small intestinal emptying upon ingestion of oral dosage forms as occurring in vivo. The gastrointestinal tract simulation system may be operated to continuously transfer, at pre-determined rates, fluids from the first compartment into said inner vessel and fluids from the said inner vessel. The skilled person will understand that a fully dynamic system may comprise relatively short interruptions that do not influence the gastrointestinal dissolution and intestinal permeation.
In embodiments according to the invention, the step of providing a dynamic gastrointestinal tract simulation system comprises the substeps of providing an empty gastrointestinal tract simulation system comprising at least two consecutive empty compartments, and filling said empty compartments with fluids simulating the physiological fluids of the gastrointestinal tract. In particular, providing a first empty compartment and filling said empty compartments with simulated gastric fluids and providing a second empty compartment, comprising an inner vessel mounted into an outer vessel, and filling said empty inner vessel with simulated small intestinal fluids and filling said outer vessel with sink solution. Said simulated gastric fluid comprising an aqueous solution. In a more specific embodiment comprising an aqueous solution buffered at a specific pH range. In a more specific embodiment comprising pepsin as a proteolytic enzyme at a specified range. In a more specific embodiment comprising gastric lipase at a certain concentration range. In a more specific embodiment, the simulated gastric fluid contains food-derived compounds at a certain concentration range. Said simulated small intestinal fluid comprising an aqueous solution. In a more specific embodiment comprising an aqueous solution buffered at a specific pH range. In a more specific embodiment comprising bile salts and/or phospholipids at a certain concentration range. In a more specific embodiment comprising proteolytic enzymes like trypsin and/or chymotrypsin at a specified range. In a more specific embodiment comprising pancreatic lipase at a certain concentration range. In a more specific embodiment comprising pancreatic amylase at a certain concentration range. In a more specific embodiment, the simulated small intestinal fluid contains food-derived compounds at a certain concentration range. Said sink solution comprising an aqueous solution that functions as the acceptor solution of the permeation device namely the solution will accept the substances that permeate out of said inner vessel through the semi-permeable membrane into said outer vessel. In a more specific embodiment comprising an aqueous solution buffered at a specific pH range. In a more specific embodiment, a solution having a specific composition allowing to dissolve a substantial high amount of the substance tested thereby assuring that the sink solution will never get saturated with the test substances during the experimental run. In a more specific embodiment, a solution comprising surfactants (such as among other TPGS, Tween80, or SDS) above their critical micellar concentration thereby solubilizing the permeated drug in said outer vessel and preventing the substance from diffusion back into said inner vessel due to micellar entrapment of the drug in said outer vessel into surfactant micelles. In an embodiment, the pH of the simulated gastric fluid can be varied over time through active control of the pH in the first vessel through pH monitoring and pH setpoint adjustments using acid and base additions. The composition, comprising concentrations of ionic species, enzymes and food compounds, of the simulated gastric fluid can be varied over time through the dynamic interplay between gastric secretions into the first vessel and gastric emptying out of the first vessel. Next to this the pH of the simulated small intestinal fluid can be varied over time through active control of the pH. The composition, comprising concentrations of ionic species, food compounds, bile salts, and/or enzymes can be varied over time through the dynamic interplay between gastric emptying from the first compartment into the inner vessel of the second compartment, duodenal secretions into the inner vessel of the second compartment and small intestinal emptying out of the inner vessel of the second compartment.
According other embodiments, the present invention relates to the method as described above, wherein the gastrointestinal tract simulation system further comprises at least one stirring system for stirring the contents of the first compartment and at least one stirring system for stirring the content of the inner and/or outer vessel. For the first compartment, the stirring system preferably comprising a magnetic drive mounted underneath the respective vessel and provided for generating a rotating magnetic field, a permanent magnet located at the bottom of the respective vessel and provided for rotating with and amplifying said rotating magnetic field. For the compartments containing both an inner and outer vessel (small intestinal compartments) the stirring system preferably comprising a magnetic drive mounted underneath the respective outer vessel and provided for generating a rotating magnetic field, a permanent magnet located at the bottom of the outer vessel and provided for rotating with and amplifying said rotating magnetic field and a permanent magnet located at the bottom of the inner vessel and provided for rotating with and amplifying the magnetic field generated by the permeant magnet located at the bottom of the outer vessel. The gastrointestinal tract simulation system is further operated to mix the contents of the inner vessel and/or the contents of the outer vessel by stirring the contents of the respective vessel.
In a second aspect, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal permeation of a substance, wherein said substance is selected from the group consisting of drugs classified according to the Biopharmaceutics Classification System as belonging to Class I, Class II, Class III, or Class IV
Preferably, the present invention relates to the method as described above, wherein said substance is primarily absorbed through passive transcellular diffusion. More specifically said substance is selected from the group of pharmaceutical compounds belonging to Class I and Class II.
In a third aspect, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal permeation of a substance, wherein the gastrointestinal tract simulation system is further operated to transfer, at pre-determined rates:
According to further embodiments, the present invention relates to the method as described above, wherein the gastrointestinal tract simulation system is further operated to transfer, at pre-determined rates: 0%-50% fresh sink solution/min fresh sink solution from a fresh sink reservoir to the outer vessel of the second compartment, and 0%-50% waste sink solution/min waste sink solution from the outer vessel of the second compartment to a waste sink reservoir. The term “fresh sink solution” as used here is to designate the sink solution, freshly prepared, without containing any permeated drug. The term “waste sink solution” as used here is to designate the sink solution that is present in the outer vessel and contains the test substance that permeated from the inner vessel into the outer vessel.
In a fourth aspect, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal permeation of a substance, the gastrointestinal tract simulation system comprises at least three consecutive compartments of which a third compartment preferably simulates the jejunum.
According to further embodiments, the present invention relates to the method as described above, wherein the gastrointestinal tract simulation system is further operated to transfer, at pre-determined rates:
In a fifth aspect, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal permeation of a substance, the gastrointestinal tract simulation system comprises at least four consecutive compartments of which a fourth compartment preferably simulates the ileum.
According to further embodiments, the present invention relates to the method as described above, wherein the gastrointestinal tract simulation system is further operated to transfer, at pre-determined rates:
In embodiments, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal absorption of a substance, further comprising operating the gastrointestinal tract simulation system so as to control, in each of the compartments, one or more or all of the following parameters: liquid flow, temperature, pH, ionic strength, head space, stirring, pressure, liquid volume, dissolved oxygen, redox potential, each in accordance with predetermined values, ranges or trajectories.
In embodiments, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal absorption of a substance, wherein a plurality of said gastrointestinal tract simulation systems are operated simultaneously and identically, and wherein one or more of said gastrointestinal tract simulation systems contain a test sample comprising the substance and one or more of the gastrointestinal tract simulation systems do not contain a test sample.
In embodiments, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal absorption of a substance, wherein each compartment comprises a vessel having an open top surrounded by a peripheral edge portion and a lid system configured to be placed onto the peripheral edge portion and to form a tight seal between the lid system and the vessel, wherein the lid system comprises a body with a plurality of passageways extending through the body and providing access to the interior of the vessel, said plurality of passageways comprising first passageways configured for receiving fluid transfer tubes and second passageways configured for mounting at least one sensor component.
In embodiments, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a method of simulating the gastrointestinal dissolution and intestinal absorption of a substance, wherein the fluid transfer system preferably comprises a plurality of peristaltic pumps for pumping fluid via fluid transfer tubes into and from each compartment.
In a further aspect, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the present invention relates to a gastrointestinal tract simulation system for simulating the gastrointestinal dissolution and intestinal absorption of a substance, comprising at least two consecutive compartments of which a first compartment simulates (at least part of) the stomach and a second compartment simulates (at least part) of the small intestine, preferably the duodenum, said second compartment comprising an outer vessel and an inner vessel mounted inside the outer vessel, said inner vessel having a wall comprising a cylindrical grid structure and a semi-permeable membrane, preferably a dialysis membrane, mounted around said cylindrical grid structure.
In embodiments according to the invention, which can occur in combination with the other aspects and embodiments of the invention which are described herein, said cylindrical grid structure has an interior volume of at least 25 ml, preferably at least 30 ml, more preferably at least 40 ml, even more preferably at least 50 ml. In embodiments according to the invention, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the ratio of the surface area of the cylindrical grid structure to the interior volume ranges from 1 to 3 cm2 per ml, preferably ranges from 1.5 to 2.5 cm2 per ml, more preferably is around 2.0 cm2 per ml. In embodiments according to the invention, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the cylindrical grid structure may have an open area ratio of at least 50%, preferably between 60% and 80%. Preferably the cylindrical grid structure has an open top and a partially closed bottom to receive the stirring magnet. In embodiments according to the invention, which can occur in combination with the other aspects and embodiments of the invention which are described herein, the ratio of the volume of the outer vessel to the interior volume is at least 3, preferably between 5 and 15, more preferably is around 10.
The invention will be explained in more detail below with reference to drawings in which illustrative embodiments thereof are shown. They are intended exclusively for illustrative purposes and not to restrict the inventive concept, which is defined by the appended claims.
The present disclosure will be described with respect to particular embodiments and with reference to certain drawings but the disclosure is not limited thereto but only by the claims. The drawings described are only schematic and are non-limiting. In the drawings, the size of some of the elements may be exaggerated and not drawn on scale for illustrative purposes. The dimensions and the relative dimensions do not necessarily correspond to actual reductions to practice of the disclosure.
Furthermore, the terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order. The terms are interchangeable under appropriate circumstances and the embodiments of the disclosure can operate in other sequences than described or illustrated herein.
Moreover, the terms top, bottom, over, under and the like in the description and the claims are used for descriptive purposes and not necessarily for describing relative positions. The terms so used are interchangeable under appropriate circumstances and the embodiments of the invention described herein can operate in other orientations than described or illustrated herein.
Furthermore, the various embodiments, although referred to as “preferred” are to be construed as exemplary manners in which the disclosure may be implemented rather than as limiting the scope of the disclosure.
The term “comprising”, used in the claims, should not be interpreted as being restricted to the elements or steps listed thereafter; it does not exclude other elements or steps. It needs to be interpreted as specifying the presence of the stated features, integers, steps or components as referred to, but does not preclude the presence or addition of one or more other features, integers, steps or components, or groups thereof. Thus, the scope of the expression “a device comprising A and B” should not be limited to devices consisting only of components A and B, rather with respect to the present disclosure, the only enumerated components of the device are A and B, and further the claim should be interpreted as including equivalents of those components.
Different aspects of the present disclosure will be described more fully hereinafter with reference to the enclosed drawings. The embodiments disclosed herein can, however, be realized in many different forms and should not be construed as being limited to the aspects set forth herein.
The gastrointestinal tract simulation system comprises at least two consecutive compartments of which a first compartment 100 simulates at least part of the stomach and a second compartment 200 simulates at least part of the duodenum, said second compartment comprising an outer vessel 220 and an inner vessel 210 mounted inside the outer vessel, said inner vessel having a wall comprising a grid structure, preferably cylindrical and a semi-permeable membrane, preferably a dialysis membrane mounted around said cylindrical grid structure. The first compartment 100 is adapted to receive to simulated gastric fluids, in particular gastric secretion 601a, and pharmaceutical substances 600a according to respectively a first volume Vfast and a second volume Vdose. In this way, the inner vessel can be used to mimic the absorption of compounds in the intestines, with the membrane mimicking the blood-intestine barrier. For example, as shown in
The grid structure 210 may for example be a 3D printed part, for example in polytetrafluoroethylene, PTFE, polyamide, polypropylene, polycarbonate, acrylonitrile butadiene styrene, polylactic acid or another material suitable for 3D printing. Preferably, the grid structure 210 is a CNC machined part, for example in a solid metal (e.g. steel, aluminium, titanium, . . . ), a plastic (e.g. PTFE, polycarbonate, polyether ether ketone) or polyoxymethylene. The outer vessel 220 is preferably manufactured in a transparent material to enable visual inspection of the contents of the compartment.
In embodiments, the grid structure may have an interior volume of at least 25 ml, preferably at least 30 ml, more preferably at least 40 ml, even more preferably at least 50 ml. For example, the ratio of the surface area of the grid structure to the interior volume ranges from 1 to 3 cm2 per ml, preferably ranges from 1.5 to 2.5 cm2 per ml, more preferably is around 2.0 cm2 per ml. In particular, the grid structure may have an open area ratio of at least 50%, preferably between 60% and 80%. Furthermore, the ratio of the volume of the outer vessel to the interior volume may be at least 3, preferably between 5 and 15, more preferably is around 10. Furthermore, the geometry of the grid structure allows to insert a pH sensor making it possible to measure and actively control the pH value of the interior volume of the grid structure.
In embodiments, as shown in
Another reason is the relatively high cylindrical liquid column both in the outer and especially inner vessels, compared to their diameter. As such, the stirring magnet needs to create a bigger vortex to also reach the top layers of the liquid column. For the outer vessel, this is less of an issue due to the space available allowing use of a large magnet, however in the inner vessel, space is a lot more limited, meaning only small magnets fit, which create only a small vortex, especially on low speeds. Furthermore, the stirring of the outer and inner vessels of the different compartments is necessary to allow active measurement and control of certain parameters, for example pH (see below), in said vessel.
By the provision of this magnetically driven stirring element, the need for a mechanical coupling of the stirring element through the wall of the vessel or the lid can be avoided, and as a result an air-tight sealing of the interior of the compartment can be better assured. The provision of the permanent magnet which rotates along and amplifies the generated magnetic field can ensure a good operation of the stirring element in the vessel.
In preferred embodiments, the permanent magnets 140, 240, 340, 440, 230, 330, 430 may be generally triangular prism-shaped or cylindrical-shaped. They may be freely moving elements in the respective vessel or be held in position by means of an additional positioning element (not shown).
Each compartment may comprise a vessel having an open top surrounded by a peripheral edge portion and an air-tight lid system configured to be placed onto the peripheral edge portion and to form an air-tight seal between the lid system and the vessel. This means that the lid system is configured to fit on the peripheral edge portion in such a way that, following a step of fixing the lid system on said edge portion, an air-tight seal is obtained. This can be achieved in many ways, for example involving one or more sealing rings, which are known per se to the person skilled in the art, and therefore need not be further described herein.
The lid system, as shown in
The method may comprise a first step S1 of sterilizing the interior of the gastrointestinal tract simulation system by introducing H2O2 gas into the empty compartments and/or by increasing the interior temperature of the compartments to a value within the range of 40° C. to 121° C. (15 psi) for a period of time within the range of 30 minutes.
The method further comprises a second step S2 of introducing a substance in the first compartment, wherein said substance is at least partially made of the compound for which the dissolution and permeation is tested.
The method further comprises operating the gastrointestinal tract simulation system to perform: a third step S3 of ‘gastric emptying’ by transferring a mixture of simulated gastric fluids and the test substance from the first compartment into the inner vessel, a fourth step S4 of ensuring the flow of the dissolved substance from said inner vessel by permeation through the semi-permeable membrane, preferably a dialysis membrane, e.g. regenerated cellulose, into the outer vessel and a fifth step S5 of ‘duodenal emptying’ by transferring simulated duodenal fluid out of the inner vessel of the second compartment. The method may further comprise of a multitude of additional steps (not shown) that are similar to S4 and/or S5, but pertain to subsequent compartments and/or vessels.
The gastrointestinal tract simulation system may be further operated to transfer (
In embodiments, a plurality of said gastrointestinal tract simulation systems are operated simultaneously and identically, and wherein one or more of said gastrointestinal tract simulation systems contain a substance and one or more of the gastrointestinal tract simulation systems do not contain a test sample.
At the start of the first experimental run, 20 mL of Sporanox® solution (corresponding to a dose of 200 mg itraconazole) was added to the stomach (without water) (Vdose 1=20 mL) whereas at the start of the second run, 20 mL of Sporanox solution was added together with 240 mL of water to the first compartment (with water) (Vdose 2=260 mL). During the experimental run the dynamics of the GIT were simulated through gastric secretion (flow rate F1=2.5 mL/min) of simulated gastric fluid into the stomach, gastric emptying (flow rate F2; combination of mono-exponential emptying of the dose and emptying of the added gastric secretion) from stomach to the inside of the inner vessel, duodenal secretion (flow rate F3=2.5 mL/min) from an exterior reservoir inside the inner vessel and duodenal emptying (flow rate F4=F2+F3) out of the inner vessel. As such, the volume of the stomach decreased monoexponentially in function of time whereas the volume of the inner vessel and the outer vessel remained constant. The pH of the content of the stomach was automatically controlled at a setpoint of pH 1.6. The pH of the duodenum (inner vessel) was kept constant at a value of 6.5 through active pH control and the secretion of duodenal fluids in the inner vessel. Active pH control was performed on the duodenal secretion, making it possible to change the buffering strength of the duodenal secretions in function of time.
The content of the stomach reactor was homogenized through stirring at 300 rpm. The content of the inner and outer vessel was homogenized through stirring at 500 rpm. The temperature of the respective vessels was controlled at 37° C.
Each experimental run was followed for 180 min and samples were taken from the first compartment, the inner vessel and the outer vessel after 7, 15, 30, 45, 60, 75, 90, 105, 120, 135, 150, 165, and 180 min. In the samples taken from the stomach and duodenum the total and solute concentration of itraconazole was determined. In the samples taken from the sink (outer vessel) the solute concentration of itraconazole was determined. Itraconazole concentrations were determined through UHPLC chromatography coupled to UV detection. All experiments were performed in biological triplicate.
In the experiment, the capacity of the experimental setup to replicate the in vivo observed complex gastrointestinal behavior of itraconazole solution is studied. Itraconazole is a weak basic drug (pKa 2 and 3.7) with a very low aqueous solubility thereby resulting in low small intestinal solute concentrations of the drug and hence low systemic bioavailability. To increase its solubility in the duodenum itraconazole is commercially available as a cyclodextrin-based formulation (Sporanox®, Janssen, Beerse, Belgium). In this formulation itraconazole is solubilized by means of 40% 2-hydroxypropyl-β-cyclodextrin.
The result show that ingestion of 20 mL Sporanox solution without the concomitant administration of water resulted in high total and solute concentrations of itraconazole in the stomach. Throughout the experiment, itraconazole was completely dissolved in the stomach as was demonstrated by the equal concentrations of total and solute drug present. The complete dissolution of itraconazole in the stomach resulted from the solubilizing effect of the cyclodextrins in combination with the low pH value of the stomach which is needed for weak basic drugs to become dissolved, as shown in
In both conditions, the concentrations of itraconazole decreased in function of time and this due to dilution with gastric secretions and removal of the compound from the stomach through gastric emptying. Calculation of the area under the curve for both the total, as shown in
Gastric emptying of the contents of the stomach into the duodenum resulted in a sharp increase in the total concentration of itraconazole in the lumen of the duodenum. Following this initial sharp increase, the total concentration of itraconazole decreased due to dilution of the content of the duodenum with duodenal secretions and removal of the drug substance through duodenal emptying. During the experiments without the addition of water, itraconazole entered the duodenum resulting in the precipitation of a fraction of the administered dose as was demonstrated by the lower concentrations of solute itraconazole as compared to the total concentration of itraconazole, as shown in
As such, it was evident that the observed similarity in stomach dilution between both conditions was not translated in the same dissolution and/or solubilization behavior in the duodenum. It can be anticipated that the lower concentrations of solute itraconazole during the experiments with water were due to the dilution of the compound in the stomach. Indeed, determination of the area under the curve of the total concentration of itraconazole in the duodenum revealed that the total concentration of itraconazole in the duodenum was decreased with 32.1% due to dilution with water, as shown in
The system according to the invention allowed to study the dynamically interconnected dissolution and permeation of itraconazole during transit through the gastro-intestinal tract. Whereas co-administration with water resulted in devastating decrease in the concentration of solute itraconazole, the driving force for permeation, the permeation data generated during the experiments revealed the absence of a substantial effect on the permeated fraction of itraconazole in between both conditions, as shown in
Hence these data suggest the existence of solubility-permeability interplay for itraconazole formulated with cyclodextrins. The increased concentration of solute itraconazole in the duodenum without water was not translated into an increased absorption of the compound indicating that the affinity of itraconazole for the cyclodextrin inclusion complexes was too high to generate substantial concentrations of free molecularly dissolved itraconazole available for permeation. Intraluminal dilution of itraconazole and cyclodextrins due to water ingestion resulted most probably in the absence of higher order cyclodextrin complexes for which the compound has high affinity, thereby decreasing the solubilization of the compound but generating the same concentrations of molecularly dissolved drug available for permeation. These data were fully in line with the data generated during the in vivo study (Berben et al., 2017).
Before the start of the fasted state experiments, 55 mL of simulated gastric juice was added to the stomach vessels, to simulate the rest volume of the stomach under fasted conditions, 30 mL of simulated duodenal juice was added to the inner vessel of the second compartment, and 300 mL of sink solution was added to the outer vessel of the second compartment. At the start of experiments, 696 mg indinavir sulfate was added to the stomach together with 250 mL of water During the experimental run the dynamics of the GIT were simulated through gastric secretion (flow rate F1) of simulated gastric fluid into the stomach, gastric emptying (flow rate F2; combination of mono-exponential emptying of the dose and emptying of the added gastric secretion) from stomach to the inside of the inner vessel, duodenal secretion (flow rate F3) from an exterior reservoir inside the inner vessel and duodenal emptying (flow rate F4=F2+F3) out of the inner vessel. As such, the volume of the stomach decreased monoexponentially in function of time whereas the volume of the inner vessel and the outer vessel remained constant. The pH of the content of the stomach was automatically controlled at a setpoint of pH 1.6. The pH of the duodenum (inner vessel) was kept constant at a value of 6.5 through active pH control and the secretion of duodenal fluids in the inner vessel. Active pH control was performed on the duodenal secretion, making it possible to change the buffering strength of the duodenal secretions in function of time. The pH of the outer vessel was set at a value of 6.5.
The content of the stomach reactor was homogenized through stirring at 300 rpm. The content of the inner and outer vessel was homogenized through stirring at 500 rpm. The temperature of the respective vessels was controlled at 37° C. (
Each experimental run was followed for 240 min and samples were taken from the stomach vessel (first compartment), the duodenum (inner vessel) and sink (outer vessel) after 7, 15, 30, 45, 60, 75, 90, 105, 120, 135, 150, 165, 180, 195, 210, 225, and 240 min. In the samples taken from the stomach and duodenum the total and solute concentration of indinavir was determined. In the samples taken from the sink (outer vessel) the solute concentration of indinavir was determined.
Two sets of fed state experiments were performed. Before the start of each of the experiments, the same volumes and fluids were added to the stomach vessel and inner and outer vessel of the second compartment as during the fasted state experiments. At the start of the experiments simulating high stomach pH fed conditions, 696 mg indinavir sulfate was added to the stomach together with 400 mL of the nutritional drink Ensure plus and 250 mL of water. At the start of the low stomach pH fed experiments, 696 mg indinavir sulfate was added to the stomach together with 400 mL of carbohydrate solution (same caloric content as Ensure plus) and 250 mL of water. 15 min after the start of the experiment, simulating the lag phase for gastric emptying under fed state conditions, the dynamics of the fed state upper GIT were simulated through gastric secretion (flow rate F1) of simulated gastric fluids into the stomach, gastric emptying (flow rate F2; combination of emptying of the added gastric secretion and the flow rate needed to empty the ingested stomach fluids within 240 min) from the stomach to the inside of the inner vessel, duodenal secretion (constant flow rate F3 which is a combination of the dynamically varying flow rates of buffer solution 606 with flow rate F3-1 and simulated duodenal fluid 602 with flow rate F3-2, which are pumped from exterior reservoirs into the inner vessel). The ratios of the flow rates of F3-1 and F3-2 were selected to generate the dynamic bile salts, phospholipid, and enzyme concentrations in the duodenum as observed in vivo. Finally, the duodenum was emptied with a constant flow rate F6 (sum of gastric emptying and duodenal secretions). As such, the volume of the stomach emptied linearly in function of time whereas the volume of the inner and outer vessel remained constant. During the high stomach pH experiments the pH value of the stomach was controlled online and was subjected to a sigmoidal decrease from an initial value of 4.6 till a final value of 1.6 over 240 min, simulating the impact of a protein rich meal on the stomach pH. During the experiments under low stomach pH fed conditions the pH of the stomach was kept constant at a value of 1.6. During each fed state experiment the pH of the duodenum (inner vessel) was kept constant at 5.8 through a combination of active pH control and buffer secretion, and the pH of the sink (outer vessel) was set at a value of 5.8 (
The content of the stomach reactor was homogenized through stirring at 500 rpm. The content of the inner and outer vessel of the second compartment was homogenized through stirring at 500 rpm. The temperature of the respective vessels was controlled at 37° C.
Each experimental run was followed for 240 min and samples were taken from the stomach vessel (first compartment), the duodenum (inner vessel) and sink (outer vessel) after 22, 30, 45, 60, 75, 90, 105, 120, 135, 150, 165, 180, 195, 210, 225, and 240 min. In the samples taken from the stomach and duodenum the total and solute concentration of indinavir sulfate was determined. In the samples taken from the sink (outer vessel) the solute concentration of indinavir sulfate was determined.
All experimental runs were performed in biological triplicate and concentrations of indinavir sulfate in the different compartments of the DIAMOD were determined through UHPLC-UV.
In the experiment, the capacity of the of the experimental setup to replicate the in vivo observed complex gastrointestinal behavior of indinavir sulfate under fasted and fed state conditions and the capacity to elucidate the important factors that contribute to the in vivo observed negative food effect is studied.
Indinavir is an HIV protease inhibitor and is a weak basic drug with pKa's of 3.7 and 5.9. Due to this feature, indinavir has a pH-dependent solubility, being highly soluble under acidic conditions but very poorly soluble at neutral pH environments. Yeh et al. (1998) demonstrated that indinavir sulfate was subjected to a substantial negative food effect when the compound was ingested together with a high caloric high fat meal as compared to ingestion of the drug under fasted state conditions. The authors attributed the occurrence of this negative food effect to the increased pH of the fed stomach probably resulting in the precipitation of indinavir in the stomach thereby reducing its small intestinal solute concentration, hence reducing its systemic exposure. Next to this, slower gastric emptying under fed state conditions as compared to fasted state conditions could have contributed to the observed effect. Carver et al. (1999) tested this hypothesis by dosing indinavir sulfate to fasted state individuals and individuals having ingested different types of isocaloric meals. The meals consisted of a high protein, high fat, and high carbohydrate meal which all had a different impact on the fed stomach pH upon ingestion. Being isocaloric, each meal resulted in the same delay in gastric emptying as compared to the fasted state condition. Whereas only the protein rich diet resulted in a substantial increase of the stomach pH, the ingestion of the high lipid and high carbohydrate meal resulted in comparable gastric pH values as present under fasted state conditions. Nevertheless, dosing of indinavir sulfate together with a high protein, high fat or high carbohydrate meal resulted in a decrease in indinavir bioavailability of 68%, 34%, or 45% as compared to dosing under fasted conditions. Considering the absence of any stomach pH effect by the high fat and high carbohydrate meals, these data indicated that the negative food effect for indinavir was not solely mediated by an increased stomach pH. Another important difference between fed and fasted state conditions is the high concentration of bile salts, phospholipids, and digestion products under fed conditions. These food-induced components can solubilize poorly aqueous soluble compounds through micellar entrapment. Notwithstanding the increase in solute drug, this micellar entrapment can result in a lower bio accessible concentration of indinavir resulting in a decreased absorption of the drug.
The results show that dosing of indinavir sulfate under fasted state conditions resulted in the complete dissolution of this compound in the stomach. This could be attributed to the low pH value of 1.6 of the stomach which was well below the pKa of indinavir namely 5.9. The total and solute concentration of indinavir decreased in function of time due to intragastric dilution of the stomach with gastric secretions and the emptying of the drug from the stomach through fast mono exponential stomach emptying (
Dosing of indinavir sulfate together with water and the carbohydrate solution resulted in the complete dissolution of the drug in the stomach as was demonstrated by the equal concentrations of total and solute of indinavir in the stomach. Indeed, during this condition the pH of the stomach was at a low value of 1.6, simulating the impact on the stomach pH upon ingestion of a carbohydrate solution. As such, a completely dissolved indinavir solution was provided to the duodenum by the stomach (
Finally, comparison of the AUC for solute concentrations of indinavir in the duodenum under the different dosing regimens clearly revealed that dosing under fasted state conditions resulted in high concentrations of solute drug in the duodenum in function of time. Dosing of indinavir sulfate under fed state conditions together with the carbohydrate solution did not decrease the AUC as compared to fasted state conditions. On the contrary the AUC was increased under this dosing regimen. This was due to the absence of any substantial precipitation of indinavir in the duodenum whereas duodenal precipitation occurred during fasted state experiments. The higher concentrations of total indinavir delivered to the duodenum and the higher pH of the duodenum under fasted conditions as compared to dosing of indinavir with a carbohydrate solution (fed state conditions) resulted in a higher driving force for precipitation. Since the pH of the fed stomach in this case was the same as during the fasted state experiment, it can be concluded that increased dilution of the indinavir dose by the larger stomach volumes under fed state conditions in combination with a slower gastric emptying did not result in lower concentrations of solute indinavir in the duodenum. On the contrary, during dosing under fed state conditions with elevated stomach pH, the AUC of solute concentration in the duodenum decreased substantially indicating that the increased stomach pH under fed conditions contributes to the lower concentrations of solute indinavir in the duodenum (
The system according to the invention allowed to study the negative food effect for indinavir sulfate thereby resulting in a strong in vitro-in vivo correlation. Furthermore, the system highlighted both the role of increased gastric pH, slower gastric emptying, and micellar entrapment of indinavir sulfate under fed state conditions as compared to fasted state conditions thereby resulting into mechanistic insights into the in vivo observed negative food effect.
Before the start of the fasted state experiments, 55 mL of simulated gastric juice was added to the stomach vessels, to simulate the rest volume of the stomach under fasted conditions, 30 mL of simulated duodenal juice was added to the inner vessel of the second compartment, 300 mL sink solution was added to the outer vessel of the second compartment, 100 mL of simulated jejunal juice was added to the inner vessel of the third compartment, 600 mL sink solution was added to the outer vessel of the third compartment. At the start of the experiments 1 capsule of 200 mg microsized fenofibrate (Lipanthyl®, Abbott Products N.V., Brussels, Belgium) or 1 tablet of 145 mg nanosized fenofibrate (Lipanthylnano®, Abbott products N.V., Brussels, Belgium) was added to the stomach together with 250 mL of water. During the experimental run the dynamics of the GIT were simulated through gastric secretion (flow rate F1) of simulated gastric fluid into the stomach, gastric emptying (flow rate F2; combination of mono-exponential emptying of the dose and emptying of the added gastric secretion) from stomach to the inside of the inner vessel of the second compartment, duodenal secretion (flow rate F3) from an exterior reservoir inside the inner vessel of the second compartment and duodenal emptying (flow rate F4=F2+F3) out of the inner vessel of the second compartment inside the inner vessel of the third compartment and jejunal emptying (flow rate F5=F4) out of the inner vessel of the third compartment. As such, the volume of the stomach decreased monoexponentially in function of time whereas the volume of the inner vessel and outer vessel of the second and third compartment remained constant. The pH of the content of the stomach was automatically controlled at a setpoint of pH 1.6. The pH of the duodenum (inner vessel of second compartment) was kept constant at a value of 6.5 through active pH control and the secretion of duodenal fluids in the inner vessel of the second compartment. Active pH control was performed on the duodenal secretion, making it possible to change the buffering strength of the duodenal secretion in function of time. The pH of the outer vessel of the second compartment was set at a value of 6.5. The pH of the jejunum (inner vessel of third compartment) was kept constant at a setpoint of 7.0 through active pH control. The pH of the outer vessel of the third compartment was set at a value of 7.0.
The content of the stomach reactor, the inner and outer vessel of the second compartment, and the inner and outer vessel of the third compartment were homogenized through stirring at 450 rpm. The temperature of the respective vessels was controlled at 37° C.
Each experimental run was followed for 240 min and samples were taken from the stomach vessel (first compartment), the duodenum (inner vessel second compartment and sink (outer vessel second compartment), jejunum (inner vessel third compartment) and sink (outer vessel third compartment) after 7, 15, 22, 30, 45, 60, 75, 90, 105, 120, 135, 150, 165, 180, 195, 210, 225, and 240 min. In the samples taken from the stomach, duodenal and jejunal compartment the total and solute concentration of fenofibrate were determined. In the samples taken from the sink of the second and third compartment (outer vessel) the solute concentration of fenofibrate was determined.
In the experiment, the capacity of the experimental setup to replicate the in vivo observed gastrointestinal behavior of nano- and microsized fenofibrate is studied. Fenofibrate is a neutral lipophilic BCS class 2 compound. The extremely low aqueous solubility of fenofibrate results in low systemic exposure upon oral administration. Therefore, fenofibrate is on the market as micro- and nanoparticles namely Lipanthyl® and Lipanthylnano®, respectively. Micro- and nanonization of poorly soluble drug compounds is an enabling strategy to increase the dissolution rate of drug compounds. Since this formulation strategy does not increase the solubility of the drug but its dissolution rate these type of formulations are typically not absorbed directly from the duodenum but their absorption also occurs in the jejunum. Therefore, Lipanthyl® and Lipanthylnano® were tested in the experimental setup that consisted of three compartments namely the stomach, duodenal and jejunal compartment as depicted in
The results show that micronization (Lipanthyl®) and nanonization (Lipanthylnano®) resulted in different gastrointestinal dissolution and permeation of fenofibrate under fasted state conditions. Gastric emptying of fenofibrate after administration of Lipanthyl® or Lipanthylnano® resulted in the presence of fenofibrate in the duodenum. In the duodenum the drug started to dissolve resulting in the presence of low concentrations of solute fenofibrate. Higher solute concentrations of fenofibrate were present in the duodenum after dosing of Lipanthylnano® as compared to Lipanthl® and this throughout the experimental run, as shown in
These data were fully in line with the data generated during an in vivo study in humans that demonstrated that intake of nanosized fenofibrate under fasted state conditions generated increased duodenal solute concentrations of fenofibrate as compared to microsized fenofibrate, which was translated in an increased systemic exposure (Hens et al., Gastrointestinal behavior of nano- and microsized fenofibrate: In vivo evaluation in man and in vitro simulation by assessment of the permeation potential, European Journal of Pharmaceutical Sciences 77 (2015), pp 40-47F1).
Other alternatives and equivalent embodiments of the present invention are conceivable within the idea of the invention, as will be clear to the person skilled in the art. The scope of the invention is limited only by the appended claims.
| Number | Date | Country | Kind |
|---|---|---|---|
| 20216750.8 | Dec 2020 | EP | regional |
| 21154616.3 | Feb 2021 | EP | regional |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/EP2021/087117 | 12/21/2021 | WO |
