MEANS AND METHODS FOR IMPLANTING CELLS SECRETING A THERAPEUTIC COMPOUND

Abstract

A device for implantation into a mammalian host has a dialysis tube with a lumen and a dialysis membrane enclosing the lumen. The dialysis tube has an outer diameter of 20 m to 600 m. The dialysis membrane has a molecular weight cutoff of from 25 kDa to 150 kDa and has a material composition of at least one of the following: polyethersulfone, polyarylethersulfone, polyethylene, polytetrafluoroethylene, polyvinylidene fluoride, and polypropylene. A therapeutic composition is disposed within the lumen and has cells that secrete a therapeutic compound. A matrix material embeds the cells. The matrix material is one or more of alginate, collagen, fibrin, extracellular matrix material, synthetic hydrogel, and acrylate. Also disclosed are a method of producing a device for implantation, a port system, and a cell secreting a therapeutic compound for use in treating disease by implantation of the cells in a device for implantation.

Claims

1. A device for implantation into a mammalian host, comprising: a) a dialysis tube with a lumen and a dialysis membrane enclosing the lumen, wherein: i) the dialysis tube has an outer diameter of from 20 m to 600 m, and ii) the dialysis membrane has a molecular weight cutoff of from 25 kDa to 150 kDa and comprises at least one material selected from the group consisting of polyethersulfone, polyarylethersulfone, polyethylene, polytetrafluoroethylene, polyvinylidene fluoride, and polypropylene; and b) a therapeutic composition disposed within the lumen and comprising a plurality of cells configured for secreting a therapeutic compound, and a matrix material embedding said cells, wherein the matrix material is selected from the group consisting of alginate, collagen, fibrin, extracellular matrix material, synthetic hydrogel, and acrylate.

2. The device of claim 1, wherein the dialysis tube has an outer diameter of from 400 m to 500 m.

3. The device of claim 1, wherein the dialysis membrane has a molecular weight cutoff of from 80 kDa to 100 kDa.

4. The device of claim 1, further comprising an implantable first guiding system configured for at least partially guiding the dialysis tube through a subcutaneous tissue.

5. The device of claim 1, further comprising a port providing access to the dialysis tubing from a body surface of the mammalian host and/or from a subcutaneous access element.

6. The device of claim 5, wherein the port is configured for transcutaneous implantation.

7. The device of claim 5, wherein the port comprises a fixing element configured to fix the port within or under the skin of the mammalian host.

8. The device of claim 1, wherein the dialysis tube is configured to be at least partially implanted intraperitoneally, subcutaneously, intraarterially, or intravenously.

9. The device of claim 1, wherein the device is configured for in situ replacement of the dialysis tube and/or for in situ replacement of the content of the dialysis tube.

10. The device of claim 1, wherein the therapeutic compound is selected from the group consisting of insulin, glucagon, Factor IX, human growth hormone (huGH), Calcitonin, Glucagon-like peptide (GLP-1) and agonists, Amylin, levodopa, somatostatin, alpha1-antitrypsin (AAT), interleukins, and interferons.

11. The device of claim 1, wherein the therapeutic compound is insulin or an insulin analog, wherein the plurality of cells is adapted to secrete an amount of insulin related to the concentration of glucose present in the matrix material.

12. A method of producing the device for implantation according to claim 1, comprising: a) filling the dialysis tube with a plurality of cells secreting a therapeutic compound and a matrix material; b) at least temporarily sealing at least one end of the dialysis tube; and c) thereby producing the device for implantation.

13. A port system, comprising: a device for implantation comprising a port according to claim 5; and an extracorporeal connecting head.

14. A cell secreting a therapeutic compound for use in treating disease by implantation of said cells in a device for implantation according to claim 1.

15. A cell secreting a therapeutic compound for use in treating diabetes, Hemophilia B, huGH deficiency, osteoporosis, Parkinson's disease, acromegaly, an endocrine tumor, alpha1-antitrypsin-deficiency, an infectious disease, an autoimmune disease, cancer, or chronic virus infection by implantation of said cells in a device for implantation according to claim 1.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0118] The above-mentioned aspects of exemplary embodiments will become more apparent and will be better understood by reference to the following description of the embodiments taken in conjunction with the accompanying drawings, wherein:

[0119] FIG. 1 is a schematic partial view of a device for implantation 10 comprising a dialysis tube 20 and a sealing means 45;

[0120] FIG. 2 is a schematic in situ view of a transcutaneous embodiment of the device for implantation 10;

[0121] FIG. 3 is a port system 200 with first guiding system 140;

[0122] FIG. 4 is a schematic view of a port system 200 comprising a first guiding system 140 with openings 170;

[0123] FIG. 5 is a schematic view of a port system 200 with sensor element 180;

[0124] FIG. 6 is a schematic in situ view of a transcutaneous embodiment of the device for implantation 10 containing a multitude of dialysis tubes 20;

[0125] FIG. 7A is an aggregation of insulin, measured as mean fluorescence intensity (MFI) in an Thioflavin T-assay, induced by membranes made of Polyvinylidene fluoride (PVDF), polyethersulfone (PES), and polyarylethersulfone (PAES);

[0126] FIG. 7B is an aggregation of insulin, measured as mean fluorescence intensity (MFI) in an Thioflavin T-assay, induced by membranes made of polytetrafluoroethylene (PTFE); and

[0127] FIG. 7C is an aggregation of insulin, measured as mean fluorescence intensity (MFI) in an Thioflavin T-assay, induced by membranes made of Polypropylene (PP), polyamide (PA), polyethylene (PE) and polyurethane (PU).

DESCRIPTION

[0128] The embodiments described below are not intended to be exhaustive or to limit the invention to the precise forms disclosed in the following detailed description. Rather, the embodiments are chosen and described so that others skilled in the art may appreciate and understand the principles and practices of this disclosure.

Example 1: Device for Implantation with Optional Port

1.1 Device for Implantation

[0129] As shown in FIG. 1, the device for implantation 10 may comprise a dialysis tube 20 and a sealing means (also referred to as a seal) 45; as shown in FIG. 1, the device for implantation is open at the bottom part, e.g., in a state immediately filling and before sealing and/or integration into a port 70. The dialysis tube has a dialysis membrane 40 enclosing a lumen 30. Disposed within the lumen 30 are at least cells secreting a therapeutic compound 50 and a matrix material 60.

1.2 Transcutaneous Embodiment of the Device for Implantation

[0130] As shown in FIG. 2, the device for implantation 10 may be adapted for transcutaneous implantation. In accordance, the device for implantation 10 may comprise the dialysis tube 20 connected to a port 70 having a first connecting element 90, in turn connected to first fixing element 80, and transcutaneous portion 130. The transcutaneous portion 130 spans the skin 100 of the mammalian host. The first fixing element 80, located within the subcutaneous tissue, fixes the port 70 in the skin 100 of the mammalian host and, together with transcutaneous portion 130 provides access to the first connecting element and the dialysis tube 20 from the exterior of the body of the mammalian host. The port may provide fluid access to the lumen 30 of dialysis tube 20, as shown in FIG. 2; it is, however, also envisaged that dialysis tube 20 is sealed at both ends and that access via the port allows removal of the dialysis tube 20 and, in an embodiment, re-introduction of a fresh dialysis tube 20.

1.3 Port System with First Guiding System

[0131] Shown in FIG. 3 is a port system 200 comprising a device for implantation 10 and an extracorporeal connecting head 160. The device for implantation comprises dialysis tube 20 and a port 70, wherein the port comprises a first fixing element 80, a transcutaneous portion 130, a first connecting element 90, and second connecting element 150. From first connecting element 90 protrudes first guiding system 140, which may guide dialysis tube, e.g., through subcutaneous tissue and/or may determine the spatial arrangement of dialysis tube 20. As shown in FIG. 3, first guiding system 140 only partially covers dialysis tube 20 over its longitudinal extension. It is, however, also envisaged that first guiding system 140 completely covers dialysis tube 20.

1.4 First Guiding System with Openings

[0132] As described herein above and as shown in FIG. 4, first guiding system 140 may comprise openings 170, e.g., holes, which may, e.g., be introduced by laser ablation. Openings 170 in first guiding system 140 provide for free diffusion access to dialysis tube 20. The diameter of the first guiding system 140 may be substantially larger than the diameter of dialysis tube 20, as shown in FIG. 4; in accordance, first guiding system 140 may also accommodate a multitude of dialysis tubes 20 and/or of loops of a dialysis tube 20. The diameter of first guiding system 140 may, however, also be selected such that only one dialysis tube 20 can be accommodated within said first guiding system 140.

1.5 Port System with Sensor(s)

[0133] As shown in FIG. 5, port system 200, may comprise one or more sensors 180 extending into dialysis tube 20, permitting recording of information on the state of the cells secreting a therapeutic compound comprised therein. Sensor(s) 180 or their connectors may extend through port 70 and, optionally, through extracorporeal connecting head 160, and may be adapted for connection to a signal transducer unit 190.

1.6 Shown Schematically in FIG. 6 is a Transcutaneous Embodiment of the Device for Implantation 10 Containing a Plurality of Dialysis Tubes 20.

[0134] As shown in FIG. 6, the device for implantation 10 may be adapted for transcutaneous implantation. In accordance, the device for implantation 10 may comprise a plurality of dialysis tubes 20 connected to a port 70 having a first connecting element 90, in turn connected to first fixing element 80, and transcutaneous portion 130. The dialysis tubes 20 may be oriented in a radiant order and may be supported by one or more spreader elements 210, which may be configured to hold the dialysis tubes 20 in the desired orientation, e.g., by comprising a spring element. The transcutaneous portion 130 spans the skin 100 of the mammalian host. The first fixing element 80, located within the subcutaneous tissue, fixes the port 70 in the skin 100 of the mammalian host and, together with transcutaneous portion 130 provides access to the first connecting element and the dialysis tubes 20 from the exterior of the body of the mammalian host. The port may provide fluid access to the lumen 30 of dialysis tubes 20, as shown in FIG. 7; it is, however, also envisaged that dialysis tubes 20 are sealed at both ends and that access via the port allows removal of the dialysis tubes 20 and, in an embodiment, re-introduction of fresh dialysis tubes 20.

Example 2: Insulin Compatibility of Different Membrane Materials

2.1 PVDF, PES and PAES

[0135] Sixteen PES (polyethersulfone) or PAES (polyarylethersulfone) membrane pieces (hollow fibers, CMA 65) were transferred to Eppendorf-tubes. Each piece was 3.0 cm in length and had a diameter of 0.5 mm (total surface area ca. 15 cm.sup.2). Polyvinylidene fluoride (PVDF) membrane strips were transferred to an Eppendorf-tube (15 cm.sup.2 surface area). Each material was incubated with 1.5 ml insulin solution (1 mg/ml human insulin dissolved in phosphate buffered saline, PBS) at 37 C. with 300 rpm rotational agitation for 24 hours. As a positive control, insulin aggregation was induced in one sample by incubation at 70 C. for 2 h.

[0136] Thioflavin T (ThT) staining was performed to assess insulin aggregation/fibril formation. 50 l of each sample was incubated with 150 l PBS and 10 l of ThT solution (450 M) for 1 h in the dark. Mean fluorescence intensity (MFI) of the samples was analyzed using a Synergy4 plate reader (BioTek Instruments) at an excitation of =450 nm and an emission of =482 nm. Results are shown in FIG. 7A). A more than 2-fold increase of MFI relative to the non-aggregated insulin control (4 C.) indicates relevant aggregation which is undesirable. PVDF, PES and PAES did not induce insulin aggregation and were identified as insulin compatible materials.

2.2 PTFE

[0137] A 25 cm long polytetrafluoroethylene (PTFE) tubing (diameter: 0.4 mm) was connected to an Accu-Check Spirit Combo insulin pump. The pump reservoir was filled with human insulin solution (3 mg/ml, Insuman Infusat, Sanofi). Pump, connected tubing and an Eppendorf tube for sample collection were placed in an incubator at 37 C. Pump basal rate was set to 0.8 U/h (=8 l/h). Additionally, 3 boli of 7 U (=70 l) were delivered within 24 h. A total sample volume of ca. 400 l was collected in an Eppendorf tube over 24 h.

[0138] The samples collected after 24 h and after 72 h were analyzed for insulin aggregation using ThT staining. 50 l of each sample was incubated with 150 l PBS and 10 l of ThT solution (450 M) for 1 h in the dark. Mean fluorescence intensity (MFI) of the samples was analyzed using a Synergy4 plate reader (BioTek Instruments) at an excitation of =450 nm and an emission of =482 nm. Results are shown in FIG. 7B). A more than 2-fold increase of MFI relative to the non-aggregated insulin control (4 C.) indicates relevant aggregation which is undesirable. PTFE did not induce insulin aggregation and was identified as an insulin compatible material.

2.3 PA, PE, PU, and PP

[0139] Tubing made either of polyamide (PA, ca. 30 cm.sup.2), polyethylene (PE, ca. 15 cm.sup.2), polyurethane (PU, ca. 30 cm.sup.2) or polypropylene (PP, ca. 30 cm.sup.2) were connected to Accu-Check Spirit Combo insulin pumps. The diameter of all tubing was 0.4 to 0.6 mm. The pump reservoirs were filled with human insulin solution (3 mg/ml, Humalog, Eli Lilly). Pumps, connected tubing and an Eppendorf tubes for sample collection were placed in an incubator at 37 C. Pump basal rate was set to 0.5 U/h (=5 l/h). A total sample volume of ca. 120 l was collected in each Eppendorf tube over 24 h. Insulin remaining in the insulin-compatible pump reservoir at the end of the experiment served as control.

[0140] The samples collected after 24 h were analyzed for insulin aggregation using ThT staining. 50 l of each sample was incubated with 150 l PBS and 10 l of ThT solution (450 M) for 1 h in the dark. Mean fluorescence intensity (MFI) of the samples was analyzed using a Synergy4 plate reader (BioTek Instruments) at an excitation of =450 nm and an emission of =482 nm. Results are shown in FIG. 7C). A more than 2-fold increase of MFI relative to the non-aggregated insulin control (pump reservoir) indicates relevant aggregation which is undesirable. PP and PE did not induce insulin aggregation and were identified as insulin compatible materials. PA and PU showed high MFI values indicating formation of insulin aggregates. PA and PU were identified as non-compatible materials.

[0141] While exemplary embodiments have been disclosed hereinabove, the present invention is not limited to the disclosed embodiments. Instead, this application is intended to cover any variations, uses, or adaptations of this disclosure using its general principles. Further, this application is intended to cover such departures from the present disclosure as come within known or customary practice in the art to which this invention pertains and which fall within the limits of the appended claims.

LIST OF REFERENCE NUMBERS

[0142] 10 device for implantation [0143] 20 dialysis tube [0144] 30 lumen of dialysis tube [0145] 40 dialysis membrane [0146] 45 sealing means [0147] 50 cells secreting a therapeutic compound [0148] 60 matrix material [0149] 70 port [0150] 80 fixing element [0151] 90 first connecting element [0152] 100 skin of the mammalian host [0153] 110 subcutaneous tissue [0154] 120 body cavity, e.g., abdomen [0155] 130 transcutaneous portion [0156] 140 first guiding system [0157] 150 second connecting element [0158] 160 extracorporeal connecting head [0159] 170 opening (hole) in first guiding system [0160] 180 sensor element [0161] 190 signal transducer unit [0162] 200 port system [0163] 210 spreader element