EP1660626A2 - Processeur de cellules servant a traiter des maladies - Google Patents

Processeur de cellules servant a traiter des maladies

Info

Publication number
EP1660626A2
EP1660626A2 EP04764395A EP04764395A EP1660626A2 EP 1660626 A2 EP1660626 A2 EP 1660626A2 EP 04764395 A EP04764395 A EP 04764395A EP 04764395 A EP04764395 A EP 04764395A EP 1660626 A2 EP1660626 A2 EP 1660626A2
Authority
EP
European Patent Office
Prior art keywords
cells
substance
substances
cell
human
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP04764395A
Other languages
German (de)
English (en)
Inventor
Ute Steinfeld
Hyeck-Hee Lee
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Korea Institute of Science and Technology Europe Forschungs GmbH
Original Assignee
Korea Institute of Science and Technology Europe Forschungs GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Korea Institute of Science and Technology Europe Forschungs GmbH filed Critical Korea Institute of Science and Technology Europe Forschungs GmbH
Publication of EP1660626A2 publication Critical patent/EP1660626A2/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/04—Cell isolation or sorting
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/08—Chemical, biochemical or biological means, e.g. plasma jet, co-culture

Definitions

  • the present invention relates to a device, hereinafter also referred to as a medical cell processor or simply referred to as a cell processor, which modifies cellular components of human or animal blood, in particular cells of the body's immune system, such as T cells or macrophages, in such a way that the Cells after their introduction into the human or animal body have a therapeutic function, for example against cancer z. B. the liver or cancer of the brain or against other diseases.
  • the cellular components described are also referred to as cells for short.
  • the cells can also be, for example, other already differentiated, endogenous cells of the human or animal body or cells that have not yet been differentiated, such as stem cells. your.
  • the cell processor can be implanted in the human or animal body, but non-implantable configurations are also possible.
  • the therapeutic function of the modified cells introduced into the human or animal body can consist, for example, in a controlled release of active substance or in a tissue regeneration or the like.
  • the active ingredient is usually supplied to the whole human or animal body.
  • the active ingredient can, for example, be administered orally or by injection; it then distributes itself evenly throughout the human or animal organism.
  • the decisive disadvantage of previous treatment methods is that unaffected regions of the human or animal body can also be affected by the active ingredients and that only a small part of the active ingredient can be effective in the target areas. Accordingly, correspondingly high doses of active ingredient are inevitable.
  • the object of the present invention is to provide a device, for example a device which can be implanted in the human or animal body, which makes it possible to modify human or animal cells in such a way that, after they have returned to the human or animal body were introduced, targeted to desired parts of the body or cells are supplied and have a therapeutic function there.
  • a device designed in this way it is thus possible, for example, with low dosage Combat diseases in a targeted manner or build up and strengthen tissues without affecting uninvolved regions of the body.
  • a device according to the invention or the cell processor has the following components: a device for isolating cells, for example from the blood circulation or a blood sample, a cell line or cell culture (for example freshly isolated cells from patients, primary cultures, etc.), advantageously a device for Fixation of cells, a device for introducing substances, for example active substances, into or for attaching these substances to the fixed cells, and advantageously also a device for determining the concentration of the substances in or on the cells.
  • the device also has a device for introducing cells into the human or animal bloodstream.
  • the cells or the cells and the medium surrounding them are also transported between the individual devices of the cell processor, for example with the aid of
  • the individual sub-devices for manipulating the cells are advantageously designed as contactless devices as far as possible, since a mechanical contact between an immune cell and a surface is an immune reaction can trigger.
  • the cell processor is implanted in the immediate vicinity of a vein (but the processor can also be arranged or carried outside the body).
  • the bioprocessor takes blood from the blood stream. Certain blood cells, e.g. Leukocytes, selected and these cells loaded with an encapsulated drug. The cells can also be encapsulated
  • the encapsulation can e.g. consist of thermally decomposable materials, so that the medication is released by heating in the target area or target area (for example by hyperthermia therapy).
  • the encapsulation can also take place with the aid of material which can be split by the cell's own enzymes (for example detran). In this case, the medication will be released after a foreseeable period of time (equivalent to the use of unencapsulated medication).
  • the carrier cell itself can be killed by the medication after a definable period of time. This makes the cell membrane permeable to the drug.
  • the defined period of time can be selected (e.g. by the type of medication, the formulation of the medication, by the choice of cytotoxic properties and / or the concentration) so that the cell can reach its destination during this time. As a result, the medication is released at the target location.
  • the cells are released back into the bloodstream and transport the active substance to the disease site, eg tumor, in a target-specific manner.
  • the system or the cell processor is thus implanted in the body or on Body worn or placed outside the body.
  • the cell processor can be used or used as a laboratory device, for example in the laboratory.
  • the device can be designed on the one hand as a system that can be worn on the body.
  • the device can also be designed as a laboratory system or as a system to be operated in the laboratory for modifying human or animal cells, for example for therapeutic purposes.
  • a laboratory system for modifying cells for autologous cancer therapies in which, for example, immune cells are isolated from a patient's tumor, then multiplied in the laboratory and finally released back into the patient's bloodstream. After the multiplication step in the laboratory, these cells can then be loaded with the device according to the invention with nanoparticles which are coated or provided with medicaments and / or nucleic acids and / or other therapeutically active compounds.
  • Uses of the laboratory device for screening purposes for pharmaceutical active substances are also conceivable.
  • the cell processor can thus be implantable, but can also be in a non-implantable shape and / or size. However, cell handling, for example isolation, fixation, transporting or counting cells etc., can also be done by touch. An example of this is cell sorting via antibody binding.
  • the first sub-device which has a cell processor designed according to the invention, is the device for isolating the cells.
  • this insulation device consists of at least one capillary, for example of a high-polymer plastic, and of at least two electrodes arranged on this capillary or these capillaries. The cells are passed through the capillary, the capillary diameter being designed so that only individual cells can pass through the capillary.
  • the individual cell types have different electrical conductivities.
  • the cells touch the electrodes, depending on the cell type, different currents can be measured. In this way, the cells can be distinguished from one another and can be selected.
  • An insulation device designed in this way thus isolates the required cells from their surroundings, for example a blood sample, by comparing the conductivities of different cell types.
  • at least one laser detector is arranged on the capillary or capillaries. Since the different cell types also have different light refraction properties, the required cells can be selected or isolated on the basis of the light refraction. In a further advantageous embodiment, the cells required are isolated by measuring the impedance, in which different cell types also differ.
  • an extension is connected at the end of at least one of the capillaries, an electromagnetic field being generated on or in this, for example with the aid of two electrodes arranged on it. Since various ne cell types have particle charges of different heights, they are influenced to different extents by the electromagnetic field and accordingly have a different duration for reaching the capillary wall in the expanded area in the electromagnetic field. In this way, the required cells can be isolated using their particle charge.
  • the electrophoretic mobility of the cells is used. This electrophoretic mobility takes on different values depending on cell properties such as density of the surface charge, volume and weight and can therefore also be used to select the desired cell type.
  • a further advantageous embodiment of the device for cell isolation uses the different particle sizes of different cell types.
  • a filter membrane is used, which is designed in such a way that only certain blood cells can pass through the membrane, other blood cells are retained.
  • Membrane filters with different pore sizes can also be used.
  • a further design option for the insulation device uses so-called solution diffusion membranes.
  • a mass transfer or isolation of the desired cell type is, for example, caused or carried out in the following manner: On the primary side of a solution diffusion membrane, a solvent is used which corresponds to all ingredients or all available cell types. On the secondary side of the solution diffusion membrane, a solvent is used that is only suitable for a certain component or a certain cell type. Only the cell type to be isolated is therefore soluble in the solvent on the secondary side.
  • the on The component soluble on the secondary side of the solvent membrane or the corresponding cell type tends to diffuse through the membrane; all other components or cell types do not.
  • the required cell type can thus be isolated.
  • Other membrane types can of course also be used for cell isolation using filtration.
  • the filtration can be implemented in an H-shaped filter module, for example.
  • the cell processor according to the invention advantageously has a device for fixing the cells.
  • the fixation serves to hold the cells so that the substance or the active ingredient can be attached to them or introduced into them.
  • a first advantageous embodiment of the sub-device for cell fixation consists of a capillary, for example made of a highly polymeric plastic, an expansion at the end of this capillary and a device for holding the cell which is introduced into the expansion.
  • the device for holding the cell is, for example, a fine needle which is applied to an electric field. The device uses the particle charges of the cells.
  • an alternating electrical field is used in the present case, which alternately holds the cell and repels it, so that the cell remains in vibration and the particle charge is retained.
  • the cells to be fixed are thus held by an alternating electrical field.
  • Such a partial device for fixing the cells with the aid of an electrical field can be used, for example, in the form of a three-dimensional microelectrode system. smooth cell manipulation.
  • Such a three-dimensional microelectrode system enables cells to be fixed or held in a cage filled with a dielectric liquid.
  • the three-dimensional microelectrode system has, for example, the following components.
  • the electrode elements which can be shaft, tube or funnel-shaped or straight or in the form of a cage or a switch, are operated by alternating current or a rotating, electrical field.
  • the electrode thickness is 10 ⁇ m and the active electrode surfaces are minimized to prevent the solution from heating up.
  • the system is operated at 5 to 11 volts and at 5 to 15 MHz.
  • the channel has a flow rate of 300 ⁇ m / s.
  • a further advantageous embodiment of the partial device for fixing cells of the immune defense is carried out as follows. With the help of a fine capillary, for example, gas or a liquid is introduced into a microball, which in turn inflates it and thus simulates a foreign body. After the defense cell has enclosed the simulated foreign body and the cell is thus fixed, the active substance is applied to the cell. The gas or liquid is then released from the microball and the defense cell floats free again. This defense cell fixation through foreign body simulation thus uses the natural function of defense cells.
  • a first advantageous embodiment of the sub-device of the cell processor for introducing a substance into the cell or for arranging a substance on the cell, which substance can be an active ingredient consists in a device for generating high-voltage pulses, for example with the aid of microelectrodes that have been coated, for example, by sputtering gold. With the help of such high-voltage pulses, which can be realized, for example, in the form of a step-like potential, small reversible pores are formed in the cell envelope. The corresponding substances or active substances are then introduced into the cells through these pores. In the present embodiment, the active ingredient or substance is thus introduced by means of electroporation.
  • a further advantageous embodiment of the sub-device for introducing or arranging substances or active substances uses magnetizable or magnetized nanoparticles or small spheres which are coated with or contain the substance or the active substance.
  • nanoparticles are particles with sizes from a few nanometers to a few hundred nanometers. In the following, however, particles with sizes are also included. understood for example in the micrometer range.
  • the sub-device now contains a device for generating a magnetic field. This can, for example, contain or consist of at least one microcoil.
  • the particles or spheres are set in motion or vibrate with the aid of this magnetic field, which is advantageously an oscillating field, but static fields are also possible, and are thereby able to penetrate the cell if the magnetic field is sufficiently strong.
  • the oscillating field can be sinusoidal or sawtoothed, for example. If the particles or spheres are in simple liquids, static magnetic fields are preferably used. If they are in more complex biological media, oscillating fields are preferred.
  • An advantageous embodiment of the magnetic field-based insertion device has a reservoir that is filled with the corresponding nanoparticles or beads. A capillary with a sluice is connected to the reservoir, the sluice only allowing a precisely defined number of nanoparticles or spheres to pass through.
  • a magnet is attached below the capillary.
  • the cell to be modified is fixed between the magnet and the capillary.
  • the substance or substances or substances are applied to the corresponding nanoparticles or beads. If necessary, these are magnetized before or after. If the magnet is activated in this arrangement, the nanoparticles or spheres are drawn into the cell.
  • the decisive factors are the time and the strength of the magnetic field: the nanoparticles or beads must not pass through the cell completely, they must remain in it.
  • Fe 2 0 3 or Fe 3 for example, come as nanoparticles or spheres containing particles or paramagnetic particles in question.
  • the cells loaded with the nanoparticles in the manner described can also be used for diagnostic purposes (comparable to the use of contrast media).
  • the nanoparticles can also be coated with contrast agents or similar substances with a high atomic number.
  • the cells loaded with nanoparticles can then also help with disease detection or better detection of diseases through the physical properties, such as magnetic characteristics (magnetized nanoparticles) of the nanoparticles.
  • the cells loaded with nanoparticles can also be used for diagnosis and at the same time for medication. This is then possible due to the magnetic properties of the nanoparticles and their simultaneous coating with an active ingredient.
  • Partial device for introducing substances or active substances into the cell uses liposomes.
  • Liposomes have the ability to penetrate the cell wall of a cell, so they can be used to transfer a substance or an active ingredient into the cell.
  • the substance or active ingredient is thus first introduced into a liposome. This is advantageously done, as described above, via active substance-coated, magnetized particles.
  • the substance or active ingredient is then transferred into the cell by lipofection, ie the liposome complex fuses with the cell membrane and releases the substance or active ingredient into the cell.
  • the partial device of the cell processor for introducing a substance or an active ingredient into the cell used phagocytosis.
  • Phagocytes are phagocytes that absorb foreign substances, dissolve and destroy them through enzymes.
  • the natural function of immune cells is thus used in that the active substance or the substance is recognized as a foreign body and is enclosed by an immune cell.
  • the substance or the active ingredient will first be brought into a form which renders it indigestible for the immune cell.
  • viruses are used. These viruses can be modified HIV viruses, for example.
  • a DNA can be introduced into the cell, which causes the cell to produce a desired active substance or substance itself.
  • a very fine needle is used for microinjection. The substance or the
  • nanofibers for example made of carbon compounds, can also be used for the injection.
  • the nanofibers advantageously have a diameter of a few tens of nanometers at the tips. If these fibers are arranged at a distance from one another corresponding to the cell size, for example on a silicon chip in a two-dimensional matrix, cells separated on the chip, for example with the aid of centrifugal forces, are only pierced by one fiber, a substance or an active ingredient being injected into the cell can.
  • a first advantageous embodiment of a partial Direction for determining the concentration of a substance or an active ingredient in or on the cell uses at least one sensor for determining magnetic field strengths.
  • a magnetic field sensor when using magnetized nanoparticles or beads, the substance or active substance concentration in the cell is measured by measuring the magnetic field strength of the nanoparticles or beads.
  • the sensitivity of the sensor is advantageously carried out in accordance with the minimal substance or agent loading, the measurement resolution in accordance with a substance or. Active ingredient loading unit. If magnetized particles are used in liposomes, the sensor can, for example, also be used to determine the number of particles loaded into a liposome.
  • the sensor is a magnetoresistive sensor or an arrangement of micro-coils that inductively detect magnetic fields.
  • the magnetic field sensors work here without contact, ie the substance or active substance concentration is determined without touching the substance or the active substance or the cell coupled to magnetic materials.
  • the active substance or substance concentration is determined with the aid of a device for measuring the fluorescent light from fluorescent dyes and / or with the aid of biomarkers.
  • a fluorescent substance or a marker substance is added to the substance or the active ingredient beforehand.
  • the line Processor has a device for determining the number or for checking the number of modified cells.
  • the partial device for determining the concentration of a substance or an active substance in or on the cell and the partial device for introducing substances or active substances into the cell are accommodated in a common reaction space.
  • This reaction space advantageously has at least two supply devices such as, for example, microchannels: a supply device for supplying the cells and a supply device for supplying substances or active ingredients or also washing reagents. The washing reagents are added after the active ingredient treatment.
  • the reaction space also advantageously has elements for electrophoresis, such as funnel-shaped or shaft-shaped microelectrodes for aligning electrostatically charged cells or straight or zigzag-shaped microelectrodes for deflecting electrostatically charged cells.
  • the sub-device for determining the concentration of a substance or an active ingredient and the sub-device for introducing substances or active ingredients into the cell can, however, also be arranged in different compartments (for example chip with different reaction spaces or compartments, so that the determination of the concentration and the active ingredient is introduced in different areas of the chip).
  • Compartments or reaction spaces are areas of the device which are delimited from one another (for example by suitable wall designs) and which have different functional areas Represent or integrate units of the device (for example, mixing chamber, electroporation unit or separation unit).
  • the individual compartments or reaction spaces can then be connected to one another via suitable flow channels, so that the cells can be guided from one compartment to the other or reaction space.
  • the cell processor has a sub-device for introducing the cells into the human or animal bloodstream.
  • This can contain, for example, micropumps, microvalves, micro nozzles and / or microfilters for controlling the flow of a fluid containing cells.
  • the modified cells are introduced into the human or animal body, they are transported to a desired defined type of tissue via the bloodstream.
  • the desired defined tissue is reached here by the ability of cells to detect this tissue with the help of messenger substances which are separated from the desired tissue. In an analogous manner, it is also possible to find previously unknown, diseased tissue.
  • the cell processor according to the invention is equipped with a device for delivering substances or active substances to a defined human or animal tissue.
  • a device for generating an electromagnetic field which is used when using magnetized nanoparticles.
  • the nanoparticles are pulled out of the immune cell by a magnetic field created with the aid of the device.
  • the release of the substance can not only be done with With the aid of a magnetic field generated by the implantable microcell processor, but also by a magnetic field generated outside or minimally invasively inside the body.
  • the substance or active ingredient is thus delivered locally to the tissue as desired. Analogous to the loading of the cells, static and / or oscillating magnetic fields can be used.
  • the active ingredient can be supplied to the desired tissue in a controlled manner and thus ideally dosed.
  • the device for delivering the substances or active ingredients is a device for destroying the modified cells at the location of the desired tissue.
  • a device can be, for example, a chemical that has been added to the substance or active ingredient that dissolves the cell on the desired tissue.
  • the trigger for self-destruction can be, for example, the concentration of messenger substances that are secreted from the desired tissue.
  • Another possible embodiment of the device for delivering substances or active substances is a device for generating ultrasound fields with a field strength sufficient to destroy cells.
  • the ultrasound field for dispensing the substance can not only be generated by the implantable microprocessor itself as described, but the dispensing of the substance can also take place by means of an ultrasound field generated outside the body or minimally invasively inside the body.
  • the cell processor according to the invention is equipped with a device for localizing modified cells, for example in the human body.
  • the localization device can, for example, be a Sensor for detecting a magnetic field caused by modified cells or a detection device for biomarkers or a detection device for fluorescent light.
  • the cell processor according to the invention relates to the integration or arrangement of a battery for energy supply. If the cell processor is used outside the body, it is advantageously possible to use a conventional battery supply. If the cell processor is used inside the body, it is advantageous to use a long-life battery, such as is also used in cardiac pacemakers (lithium-iodine batteries). Such a long-life battery can be replaced with a minimally invasive procedure.
  • the cell processor according to the invention is equipped with a contactless, inductive power supply.
  • a first coil with rectifier is used at the location of the implanted cell processor, which feeds an accumulator or a capacitor, such as a scap, which in turn represents the energy supply of the cell processor.
  • a scap is a powerful double-layer capacitor, in which the electrical energy is stored by shifting the charge at the interface between the electrode - usually made of carbon - and the organic electrolyte.
  • the capacitor or the accumulator is charged inductively via the first coil by a second coil which is applied to the body surface and which is applied to an alternating field.
  • This outer second coil can, for example, be fastened to the body by a band in the sleep phase.
  • the power supply is provided by using special carbon nanotubes. These carbon nanotubes generate an electrical charge when flowing through a liquid, for example blood. The required energy is thus made available by the liquid flow.
  • At least one reservoir is provided on or on the cell processor.
  • Such a reservoir is used to hold at least one substance or active ingredient.
  • the substances or active ingredients can be, for example, therapeutic agents such as medicines, medication precursors (prodrugs),
  • Hormones enzymes for the cleavage of drug precursors, viruses, which are used for example for gene therapy, or nanoparticles. If the reservoirs are exhausted, they can, for example, be implanted from the outside by a minimally invasive procedure, e.g. can be refilled with a fine needle. The filling can be implemented, for example, using various septa.
  • Another advantageous embodiment of the cell processor according to the invention relates to the integration of a so-called home monitoring system, which uses a transmitter to report the need for replenishment of substances or active ingredients.
  • the cell processor according to the invention or parts thereof consist of biocompatible material and / or different types of metal, such as, for example, silver, titanium or V2A, and / or of ceramic and / or plastics, such as, for example, polyethylene, silicone, polymer 908 the surface of the cell processor partly modified in such a way that it is not recognized by the immune system as a foreign body, or the cell processor secretes substances that suppress local defense reactions of the body (eg steroids).
  • metal such as, for example, silver, titanium or V2A
  • ceramic and / or plastics such as, for example, polyethylene, silicone, polymer 908 the surface of the cell processor partly modified in such a way that it is not recognized by the immune system as a foreign body, or the cell processor secretes substances that suppress local defense reactions of the body (eg steroids).
  • the cell processor described above for modifying cells is distinguished by a number of significant advantages. It enables the targeted transportation of active ingredients of various types
  • T-lymphocytes T-lymphocytes
  • monocytes neutrophils
  • neutrophils the latter after stimulation, for example with ⁇ -glucan; see Hong et al., '2003, Cancer Research 63, 9023-9031
  • other blood cells such as T-lymphocytes, monocytes or neutrophils
  • Blood cells are detected by the cell processor and substances, active substances or therapeutic agents, such as medicines, prodrugs, hormones, enzymes for cleaving the prodrugs, viruses, e.g. used for gene therapy, or nanoparticles, transferred to it.
  • the defense cells modified in this way are returned to the human or animal bloodstream.
  • the body's immune system has the natural ability to recognize certain tissues.
  • Targeted treatment measures can also be used to make a tissue recognizable for the immune system, i.e. targeted immune cells are directed to a specific tissue and the recognition of this target tissue is reinforced by immune cells.
  • An example is the targeted heating of a tumor, e.g. through hyperthermia or thermotherapy.
  • So-called heat shock proteins such as HSP 96, HSP 72, are formed in the heated tissue. These can play a role in the complex process leading to antigen presentation on the cell surface. Furthermore, they can also be released into the extracellular environment for the immune system to serve as a sign of abnormal, dead or damaged cells. An immune response in this tissue is strengthened.
  • Another example is immunotherapy with specific or multi-tumor-specific epitopes or induction of the immune response by infiltration of appropriate antigen-presenting cells or effector cells of the immune system, which likewise increase the strengthening of the immune system in the tumor tissue.
  • DNA vaccination is also conceivable.
  • the medication transport through loaded immune cells becomes more specific and effective.
  • the modified cells can thus be used to target specific tissue using the natural ability of the body's immune defense and to release the active substance at the desired position.
  • the type of transfer of the active ingredient to the transport medium with the aid of the cell processor according to the invention can vary as described. It For example, both an adsorption of the active substance on the surface and an incorporation into the transport medium are possible.
  • the type of active ingredient accumulation can also vary as described. Mechanisms such as electroporation, lipofection or microinjection are possible.
  • the cell processor according to the invention can transfer the active ingredients to the transport medium both inside the body and outside the body.
  • the medication or other components can be transported through the immune cells by storing the substance to be transported in the immune cell, or in a further envelope, such as e.g. a liposome, or is coupled to its surface (in the latter case, therefore, the substance itself is not directly embedded in the cell, but encapsulated in the further envelope, incorporated in the cell or introduced into the cell attached to the surface of this envelope).
  • a further envelope such as e.g. a liposome
  • the therapy according to the invention can be combined with all other therapies, e.g. Cancer vaccination, strengthening of the body's immune defense by e.g. Cytokine administration, etc. can be combined.
  • the therapies can be combined independently of one another, but they can also be used in parallel, as a previous therapy or as a subsequent therapy to other therapies.
  • the therapy according to the invention is combined in the manner with other therapies, that for example first with other therapies, such as, for example, by strengthening the body's immune defense or cancer vaccination, the cells required for the intended purpose are increased first and then for the therapy according to the invention can be separated from the bloodstream.
  • medications are also transported in the therapy according to the invention which are coupled to an antibody which recognizes the cell to be treated.
  • An antibody can be coupled to the body's own defense cell, which specifically docks the defense cell to the cell to be treated.
  • RNA e.g. siRNA
  • DNA are transported, which are linked to the cell to be treated or its genetic information, whereby the cell e.g. is made harmless, is prevented from further growth or cell division, or is made more recognizable by the immune system and, as a result, cells of the immune system specifically target the treated cells (for example cancer cells).
  • RNA e.g. siRNA
  • DNA are transported, which are linked to the cell to be treated or its genetic information, whereby the cell e.g. is made harmless, is prevented from further growth or cell division, or is made more recognizable by the immune system and, as a result, cells of the immune system specifically target the treated cells (for example cancer cells).
  • Cell processors according to the invention for modifying human or animal cells can be designed as described in one of the following examples.
  • FIG. 1 shows schematically an inventive Cell processor
  • FIG. 2 shows a three-dimensional view of the same
  • FIG. 3 shows devices for loading active substances and for measuring active substance concentration in a common reaction space
  • FIG. 4 shows a partial device of the cell processor for cell isolation by measuring the conductivity
  • FIG. 5 shows a partial device for cell isolation by means of a laser detector
  • FIG. 6 shows a partial device for cell isolation by means of the particle charge
  • Figure 7 shows part of devices for cell isolation using filters
  • Figure 8 shows a part of apparatus for cell fixation by means of an alternating field
  • Figure 9 shows a component device for cell fixation by means of a micro-ball
  • Figure 10 shows the electroporation of a Cell
  • FIG. 11 shows a partial device for loading cells by means of magnetized particles
  • FIG. 11 shows a partial device for loading cells by means of magnetized particles
  • FIG. 12 shows the loading of a cell by lipofection
  • FIG. 13 shows the loading of a cell by phagocytosis
  • FIG. 14 shows the loading of a cell m it with the help of a virus
  • FIG. 15 shows the loading of a cell by microinjection
  • FIG. 16 shows the discharge of the active substance from a cell with the aid of an ultrasound field
  • FIG. 17 shows an inductive power supply for the cell processor
  • FIG. 18 shows a magnetoresistive sensor and an arrangement of micro-coils
  • FIG. 19 shows the active substance loading and unloading with the aid of magnetic liposomes or magnetic nanoparticles.
  • FIG. 1 shows schematically a possible embodiment of a cell processor according to the invention.
  • Blood is supplied from a blood reservoir 1, the human blood circulation, via a filter device 3a to the cell isolation device * 4, which contains the desired, i.e. H. selected the cells to be modified.
  • a filter liquid from a filter liquid reservoir 3 can be added via a microvalve 2.
  • the amount of cells isolated is determined with the aid of a counting device 5, a device for measuring impedance.
  • the cells are fixed in a loading device 6a and loaded with the desired active ingredient.
  • the active ingredient is added via a lock 7 from an active ingredient reservoir 8.
  • the concentration of the active substance in the loaded cells is determined in the measuring device 6b.
  • the modified cells are supplied to the human bloodstream through an outlet 10.
  • the cells are transported using a micropump 9.
  • FIG. 2 shows a three-dimensional view of an embodiment of the cell processor according to the invention.
  • Blood and its cellular components flow into the microcell processor through an inlet la. about A filter structure 3 a leads the cells to the cell isolation device 4.
  • the cells With the aid of the sub-device 6a, the cells are loaded with the desired active ingredient, which is supplied with the aid of a reservoir 8.
  • the concentration of the active substance introduced is measured with the aid of the device for determining the concentration of the active substance 6b.
  • the modified cells are fed back into the bloodstream via the outlet 10.
  • the cell is transported within the system with the aid of a micropump 9, the device 3 represents a tank for a filter liquid.
  • FIG. 3 shows an integrated device 6 for fixing the cells and for loading the 1 cells with them
  • Active substances with the aid of magnetic fields generated by microcoils 6a and for measuring the concentration of the active substances in the loaded cells by means of a magnetoresistive sensor 6b.
  • FIG. 4 shows a device for isolating cells based on their conductivity.
  • the figure shows a capillary 12 with an upwardly flared end. In this widened end and in the narrow part of the capillary 12 there are cells 11.
  • FIG. 1 shows a device for isolating cells based on their conductivity.
  • FIG. 5 shows a device for cell isolation using a laser detector.
  • a capillary 12 with a funnel-shaped end and cells 11 are shown.
  • a laser detector 14a is arranged on the left narrow side of the capillary 12.
  • the light refraction properties of the cells 11 are determined with the aid of a laser beam 14b. Since different cell types differ in their refractive properties, the cell types can be distinguished with the aid of the device described. In this way, the cell types sought are isolated.
  • FIG. 6 shows a device for cell isolation based on the particle charge of the cells.
  • a capillary 12 is shown with an end that widens upwards in the shape of a funnel.
  • An extension 15 is connected to the lower end of the capillary 12.
  • Cells 11 are shown in the funnel-shaped end, in the narrow part and in the widening of the capillary 12, two electrodes 13a and 13b are sketched to the left and right of the widening. With the help of the electrodes 13a and 13b, an electromagnetic field is applied in the area of the extension 15. Due to the different particle charges, different cell types have different running times in the electromagnetic field. On the basis of this runtime, the different cell types can thus be distinguished or the desired cell type can be isolated.
  • FIGS. 7A and 7B show devices for isolating cells with the aid of filter modules.
  • FIG. 7A shows an H-shaped filter module 16a.
  • a filter membrane ran 16c introduced, which divides the H-shaped filter module 16a into two equal sections, an upper section and a lower section.
  • a solvent 16d in which all existing cell types are dissolved.
  • these are a searched cell type 11a and another cell type 11b.
  • a solvent 16b which is only suitable for the cell type 11a sought.
  • the sought-after cell type 11a thus endeavors to diffuse through the membrane 16c, but not all other cell types or components.
  • the cell isolation or selection is therefore carried out with the aid of a filter membrane 16c and two suitably selected solvents 16b and 16d.
  • Figure 7B outlines a multi-stage filter process.
  • the cells are supplied to the filter device through an inlet channel 16e.
  • Three filters 16i, 16j and 16k are integrated in the filter device. These three filters are used to separate particles with different diameters.
  • cells of a first diameter are discharged through a channel 16f
  • cells of a second diameter are discharged through a channel 16g
  • cells of a third channel 16h are discharged.
  • FIG. 8 shows a device for cell fixation by means of an alternating electrical field.
  • a capillary 12 is shown with a funnel-shaped end widening upwards and an extension 15 connected downwards.
  • Cells 11 are shown in the entire area of the capillary.
  • a needle 17a is inserted into the widening from the left. With An electric field is applied to the needle by means of a second electrode 17b. Due to the particle charge, the cells 11 can be held with the aid of the needle 17a. In order to hold the cells 11 over a longer period of time, it is necessary to maintain the particle charge. For this purpose, the cells 11 must be subjected to a certain movement. Therefore, an alternating electrical field is used in the present case; which alternately holds and repels a cell 11 so that it remains in vibration and thus the particle charge is retained.
  • FIG. 9 shows a device for fixing the cells with the aid of a microball.
  • a capillary 12 is shown, at the right end of which a microball 18a is connected.
  • the microball is inflated, • which happens with the help of a gas or a liquid 18b passed through the capillary. Inflation simulates a foreign body.
  • a defense cell 11 encloses this foreign body.
  • the defense cell 11 is thereby fixed and an active substance 19 is introduced.
  • the lower part of the picture shows in an analogous manner how the liquid or gas is removed from the microball 18d, the microball thus collapses 18c and the defense cell 11, into which the active ingredient 19 is now introduced, floats freely again ,
  • FIG. 10 shows how an active substance is introduced into a cell with the aid of high-voltage pulses, so-called electroporation.
  • FIG. 10A shows an immune cell 11, on the upper part of which reversible pores 20a are formed in the cell envelope by high-voltage pulses 20b. Through this An active substance 19 is introduced into the immune cell 11 through the pores 20a.
  • FIG. 10B outlines a corresponding device for electroporation. Two cells 11a and 11b are immobilized on a plate 20g equipped with corresponding recesses 20h. Electrodes 20c and 20d are located on the left and right edges of the depressions 20h. A voltage source 20i is applied to this, which is outlined for the cell 11a.
  • a reservoir 20f which is filled with the active substance 19 to be introduced.
  • the active substance 19 is introduced into the pores via microholes 20e located below the recesses 20h.
  • FIG. 11 shows a device for loading a cell with an active ingredient by means of magnetized nanoparticles.
  • a capillary 12 is shown with a funnel-shaped upper end.
  • the capillary 12 has a lock device 21b.
  • the funnel-shaped upper end of the capillary 12 serves as a reservoir for nanoparticles 21a.
  • An immune cell 11 is shown below the capillary, below this a magnet 21c.
  • the lock device 21b ensures that only a precisely defined number of nanoparticles 21a can pass through the lock. With the help of the magnetic field, this defined number of nanoparticles with the active substances is drawn into the immune cell 11.
  • the decisive factor in this device is the time and the strength of the magnetic field, so that the nanoparticles 21a do not completely pass through the immune cell 11, but remain in it.
  • a magnet For example, static and oscillating fields are possible.
  • suitable nanoparticles 21a are Fe 3 0 4 particles or paramagnetic particles. With the help of the particles, the active ingredient is brought into a form that makes it indigestible for the immune cell. This will be described in more detail later (relating to FIG. 13).
  • FIG. 12 shows the introduction of an active substance into an immune cell 11 using lipofection.
  • FIG. 12A shows circular liposomes 22a. These liposomes are loaded with the desired active ingredient 19 and with magnetized particles 22b.
  • An immune cell 11 is shown in the right part of FIG. 12A, inside which there are four liposomes 22a.
  • the liposomes 22a have the ability to penetrate the cell wall of an immune cell 11 and thus to transport the active substance 19 introduced into it into the cell 11.
  • the liposomes are directed in the direction of the cell 11 by the magnetized particles 22b introduced into them by means of a magnet 22c. This is shown by arrows in FIG. 12A.
  • FIG. 12A shows circular liposomes 22a. These liposomes are loaded with the desired active ingredient 19 and with magnetized particles 22b.
  • An immune cell 11 is shown in the right part of FIG. 12A, inside which there are four liposomes 22a.
  • the liposomes 22a have the ability to penetrate the cell wall
  • FIG. 12B outlines the determination of the concentration of an active substance in a cell processor according to the invention.
  • the figure shows a liposome 22a on the left, into which magnetic particles 22b and the active substance 19 have been introduced.
  • the liposome is introduced into a cell 11 by means of lipofection, which is outlined by an arrow.
  • the concentration of the active substance is determined using a magnetic sensor 22c. This is also indicated by an arrow.
  • FIG. 13 shows the loading of an immune cell 11 with an active ingredient 19 by phagocytosis.
  • An immune cell 11 and an active substance are in the upper part of the figure 19 shown.
  • the same immune cell 11 is shown in the lower area of the figure, but has completely enclosed the active substance 19.
  • the natural function of an immune cell 11 is used in phagocytosis.
  • the active substance 19 is recognized as a foreign body and enclosed by the immune cell 11.
  • the active substance 11 is previously brought into a form which renders it indigestible for the immune cell 11.
  • the indigestibility can be done, for example, by applying or enveloping the active ingredient by means of an external encapsulation or by special formulation of the active ingredient. This also protects the cell itself from the effects of the drug.
  • the encapsulation or formulation has properties that it dissolves at the target location, for example by supplying external heat to the target or by dissolving under certain physiological conditions.
  • the drug only works at the destination.
  • the drug can also be a preliminary stage (pro-drug) that is only converted into the active form at the destination.
  • FIG. 14 shows the loading of an immune cell 11 with an active substance using a modified HIV virus 23a.
  • a virus 23a is shown in the form of a cut ball. This virus 23a contains the active substance 19 in its interior.
  • An immune cell 11 is shown in the lower image area. The virus 23a adheres to the immune cell 11 with the aid of four leg-shaped projections 23b. On the underside, the virus 23a has a tube-shaped protuberance 23c which extends into the immune cell 11.
  • the virus 23a is used to introduce the active substance 19 into the immune cell 11.
  • the virus 23a can also be used to introduce a DNA into the cell 11, which causes the cell 11 to produce the corresponding active ingredient 19.
  • FIG. 15 shows the loading of a cell 11 with an active ingredient 19 by microinjection.
  • the figure shows an immune cell 11, a needle 24, and an active ingredient 19, which is partly in the needle 24 and partly already in the immune cell 11.
  • the active ingredient is injected by microinjection with a very fine needle 24 of the immune cell 11.
  • FIG. 16 shows the release of an active substance 19 on the desired tissue using ultrasound.
  • a cell 11 loaded with an active ingredient 19 is outlined on the left in the figure. After it has migrated into the desired tissue, the active substance 19 is released from the cell 11 with sufficient intensity using an ultrasound field 25; shown on the right in the picture.
  • FIG. 17 shows a device for inductive power supply for the cell processor.
  • a first coil 26b with the associated power supply 26c is sketched in a rectangular shape.
  • an implant 26d which contains a cell processor, is indicated as an ellipse.
  • the skin surface 26a is located between the first coil 26b and the implant 26d.
  • a second coil 26f, a rectifier 26g and a capacitor or accumulator 26h are drawn in the implant 26d.
  • the cell processor is supplied with energy as follows: The coil 26b is applied to an alternating field. By induction, this results in a current in the coil 26f, which is the accumulator or Capacitor 26h feeds.
  • FIG. 18 shows the determination of the concentration of an introduced active substance with the aid of a magnetoresistive sensor.
  • FIG. 18A shows an active substance-loaded liposome 22a, which contains magnetic particles, in the form of a cut-open sphere.
  • a three-dimensional view of a magnetoresistive sensor 27a is sketched below the liposome 22a. This essentially consists of three layers, the uppermost Si 3 N 4 layer 27b, the magnetoresistive film 27c underneath, and a silicon substrate 27d underneath.
  • An electromagnet 22c is drawn below the magnetoresistive sensor 27a. The position of the liposome 22a can be influenced using the electromagnet 22c.
  • the concentration of the active substance loaded into the liposome 22a is determined with the aid of the magnetic field caused by the magnetic particles by the magnetoresistive sensor 27a.
  • FIG. 18b shows an arrangement of microcoils 27e for generating or for detecting a magnetic field.
  • Figure 19 outlines the loading and unloading of a cell using magnetic fields.
  • a liposome 22a is drawn at the top left of the figure, which is loaded with an active ingredient and with magnetic particles. Some magnetic particles 21a are drawn below.
  • a magnet 21c with its magnetic field is sketched in the middle of the figure. Right in the
  • Figure is a cell 11 outlined.
  • the magnetized liposome 22a can be influenced in its position or in its path. This is shown by two arrows.
  • the magnetic nanoparticles 21a can also be supplied to the cell 11 with the aid of the magnet 21c or its magnetic field.
  • a discharge of the drug-coated particles 21a from the cell 11 is also possible. This is outlined by two arrows.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Sustainable Development (AREA)
  • Molecular Biology (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Physiology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention concerne un microprocesseur de cellules médical qui modifie des composants cellulaires du sang, en particulier des cellules du système de défense immunitaire du corps, de façon qu'après leur introduction dans le corps, les cellules exercent une fonction thérapeutique contre des maladies cancéreuses ou d'autres maladies. Le processeur de cellules selon l'invention peut être implanté dans le corps d'un être humain ou d'un animal, et comporte un dispositif pour isoler des cellules, un dispositif pour fixer des cellules, un dispositif pour introduire des substances dans les cellules, ainsi qu'un dispositif pour déterminer la concentration des substances dans les cellules. Ledit processeur de cellules peut également être utilisé à l'extérieur du corps pour charger des cellules.
EP04764395A 2003-08-29 2004-08-23 Processeur de cellules servant a traiter des maladies Withdrawn EP1660626A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10339905A DE10339905B4 (de) 2003-08-29 2003-08-29 Implantierbarer Mikrozellprozessor zur Krankheitsbehandlung
PCT/EP2004/009414 WO2005028608A2 (fr) 2003-08-29 2004-08-23 Processeur de cellules servant a traiter des maladies

Publications (1)

Publication Number Publication Date
EP1660626A2 true EP1660626A2 (fr) 2006-05-31

Family

ID=34223223

Family Applications (1)

Application Number Title Priority Date Filing Date
EP04764395A Withdrawn EP1660626A2 (fr) 2003-08-29 2004-08-23 Processeur de cellules servant a traiter des maladies

Country Status (4)

Country Link
US (1) US20070191819A1 (fr)
EP (1) EP1660626A2 (fr)
DE (2) DE10339905B4 (fr)
WO (1) WO2005028608A2 (fr)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102007010046A1 (de) * 2007-03-01 2008-09-04 Siemens Ag PCa-Nachweisgerät
EP2455774B1 (fr) * 2010-11-19 2013-08-21 ARGO-HYTOS GmbH Dispositif de capteur et son procédé de fonctionnement
CN103816578B (zh) * 2014-03-05 2016-04-27 广州一代医药科技有限公司 一种抗肿瘤磁性纳米粒子药物的靶向给药装置
DE102014104511A1 (de) * 2014-03-31 2015-10-01 Leibniz-Institut Für Analytische Wissenschaften - Isas - E.V. Verfahren und Vorrichtung zur nicht invasiven Bestimmung von Prozessparametern bei Mehrphasenströmungen
EP4496877A1 (fr) * 2022-03-23 2025-01-29 Universiteit Twente Micro-aiguille magnétique pour isoler des cellules uniques marquées par immunomagnétisme

Family Cites Families (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5128257A (en) * 1987-08-31 1992-07-07 Baer Bradford W Electroporation apparatus and process
JPH04504216A (ja) * 1989-03-31 1992-07-30 ザ・リージエンツ・オブ・ザ・ユニバーシテイ・オブ・ミシガン 細胞の特定部位点滴注入または細胞の部位特異的形質転換による疾患の治療及び該治療のためのキット
JP3203599B2 (ja) * 1989-10-24 2001-08-27 カイロン コーポレイション 感染性タンパク質デリバリーシステム
EP0488152A3 (en) * 1990-11-30 1992-11-25 Hitachi, Ltd. Method for immunoassay and apparatus therefor
JPH08511680A (ja) * 1993-03-23 1996-12-10 シービーアール・ラボラトリーズ・インコーポレーテツド 細胞内への生物学的活性物質の封入のための方法および装置
SG99279A1 (en) * 1994-12-13 2003-10-27 Peter K Law Myoblast therapy for cosmetic treatment
US5753477A (en) * 1996-03-19 1998-05-19 University Technology Corporation Magneto-biolistic methods
EP0808897A1 (fr) 1996-05-21 1997-11-26 I.D.M. Immuno-Designed Molecules Cellules présentant l'antigène, procédé de leur fabrication et leur utilisation comme des vaccins cellulaires
US6074827A (en) * 1996-07-30 2000-06-13 Aclara Biosciences, Inc. Microfluidic method for nucleic acid purification and processing
CA2383791C (fr) * 1999-09-03 2012-11-06 Miltenyi Biotec Gmbh Procedes de modification de cellules selectionnees dans une colonne de separation magnetique de cellules
US20040141974A1 (en) * 2000-02-25 2004-07-22 Boyd Robert Simon Diagnostic and therapeutic methods
CA2430554A1 (fr) * 2000-12-01 2002-07-25 Nephros Therapeutics, Inc. Dispositif de traitement sanguin intravasculaire et utilisation de ce dispositif
US7244232B2 (en) * 2001-03-07 2007-07-17 Biomed Solutions, Llc Process for identifying cancerous and/or metastatic cells of a living organism
US7288085B2 (en) * 2001-04-10 2007-10-30 Medtronic, Inc. Permanent magnet solenoid pump for an implantable therapeutic substance delivery device
WO2003045428A2 (fr) * 2001-11-30 2003-06-05 Medigene Aktiengesellschaft Utilisation d'une cellule techniquement modifiee en tant que vaccin pour traiter une tumeur
WO2003066191A1 (fr) * 2002-02-04 2003-08-14 Colorado School Of Mines Separations de particules cellulaires et colloidales par ecoulements laminaires
AU2003301322A1 (en) * 2002-10-16 2004-05-04 Yoshiro Okami Apparatus for introducing biological material, method of introducing biological material and magnetic support for introducing biological material

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO2005028608A2 *

Also Published As

Publication number Publication date
DE10339905A1 (de) 2005-03-31
DE10362104B4 (de) 2008-02-14
WO2005028608A3 (fr) 2006-01-05
DE10339905B4 (de) 2009-04-23
WO2005028608A2 (fr) 2005-03-31
US20070191819A1 (en) 2007-08-16

Similar Documents

Publication Publication Date Title
DE69921489T2 (de) Elektrotransportvorrichtung mit klingen
US20250303150A1 (en) Interventional drug delivery system and associated methods
Saltzman et al. Building drug delivery into tissue engineering design
Grayson et al. Electronic MEMS for triggered delivery
DE69835228T2 (de) Verfahren zur elektropermeabilisation einzelner zellulärer und organeller strukturen
US20240149030A1 (en) Controlled flow drug delivery implantable device
Khademhosseini et al. Drug delivery and tissue engineering
CN107988072A (zh) 具有微通道的器官模仿装置及其使用和制造方法
Li et al. Advances in magnetic microneedles: from fabrications to applications
Hassan et al. Microfluidic technologies for local drug delivery
WO2007082737A1 (fr) Système microfluidique et procédé d'exploitation
DE102004054536A1 (de) Multimodal veränderte Zellen als zellulare Darreichungsformen für aktive Substanzen und als diagnostische Zellpartikel
Prausnitz Electroporation
WO2018026925A1 (fr) Dispositif de barrière hémato-encéphalique tubulaire vasculaire
WO2005028608A2 (fr) Processeur de cellules servant a traiter des maladies
Jadeja et al. The rise of electrosomes: a revolutionary approach to nanomedicine and precision therapy
Patel et al. Nano drug delivery systems for space applications
Huang et al. A minimally invasive in vivo electroporation method utilizing flexile electrode and microneedle roller
Aceves-Serrano et al. Microfluidics for drug delivery systems
Taddie et al. The role of nanotechnology in medicine
Chetty Nanomedicine and drug delivery-revolution in health system
Khan Nano-biotechnology
Lenin et al. Nanotechnology for Sustainable Energy Storage Devices in Medical Applications
Manglani et al. Advances in Nano medicine
Raihana et al. Importance of Nanotechnology in Pharmaceutical Formulations-A Review

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20060323

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LI LU MC NL PL PT RO SE SI SK TR

AX Request for extension of the european patent

Extension state: AL HR LT LV MK

DAX Request for extension of the european patent (deleted)
17Q First examination report despatched

Effective date: 20091006

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20121005