Gene delivery by microneedle injection

• US 5,697,901 A
• Filed: 05/19/1995
• Issued: 12/16/1997
• Est. Priority Date: 12/14/1989
• Status: Expired due to Term

• Current Assignee: Applied Tissue Technologies LLC
• Original Assignee: Elof Eriksson
• Inventors: Eriksson, Elof
• Primary Examiner(s): Buiz, Michael Powell
• Assistant Examiner(s): Nguyen, A. T.
• Application Number:
  US08/445,265
• Time in Patent Office: 942 Days
• Field of Search: 604/46, 604/191, 604/890.1, 604/891.1, 604/51, 435/172.1, 435/172.2, 435/172.3, 435/240.1, 435/240.2, 435/240, 435/241
• US Class Current: 604/46
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/0021 Intradermal administration, e.g. through microneedle arrays, needleless injectors mechanical aspects A61M
• CPC Class Codes: A61K 9/1676 having a drug-free core with discrete complete coating layer containing drug adsorbates of liquid drug formulations on inert powders without simultaneous granulation step A61K9/141; with further drug-free outer coating A61K9/5078; drug conjugated to non-active particles A61K47/6921
• CPC Class Codes: A61L 2400/06 Flowable or injectable implant compositions
• CPC Class Codes: A61L 27/36 containing ingredients of undetermined constitution or reaction products thereof , e.g. transplant tissue, natural bone, extracellular matrix isolated natural compounds, e.g. collagen A61L27/24
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: A61M 2037/0023 Drug applicators using microneedles
• CPC Class Codes: A61M 2037/003 having a lumen
• CPC Class Codes: A61M 2037/0061 Methods for using microneedles
• CPC Class Codes: A61M 2205/0244 Micromachined materials, e.g. made from silicon wafers, microelectromechanical systems [MEMS] or comprising nanotechnology
• CPC Class Codes: A61M 37/0015 by using microneedles
• CPC Class Codes: A61M 5/3287 Accessories for bringing the needle into the body; Automatic needle insertion A61M5/20, A61M5/31525 take precedence
• CPC Class Codes: C12N 2510/00 Genetically modified cells

Abstract

Direct gene transfer of genetic material into an external or internal target cell site ("microseeding"), in optional combination with a wound treatment chamber, are particularly effective as a way of obtaining long term expression of native or non-native polypeptides in a host. A wide variety of proteins and materials can be expressed, either for secretion into the general blood and lymphatic system, or to alter the properties of the protein, for example, to not express proteins eliciting an immune response. The use of the optional wound chamber system for gene transfer to skin target sites also allows non-invasive assessment of the success of transfer by assaying for the presence of the expressed protein in wound fluid, in contrast to the prior art use of invasive techniques, such as biopsies, in order to achieve the same assessment of early expression.

Claims

1. A method for introducing genetic material into a cell the method comprising the step of

repeatedly injecting the genetic material into a plurality of cells of a target tissue at an in situ site on a human or non-human animal with a microneedle, whereby a change in the animal attributable to the delivery of the genetic material is detectable.

2. The method of claim 1 wherein the cell is selected from the group consisting of a superficial keratinocyte, a stem cell keratinocyte, a dermal fibroblast, a smooth muscle, a striated muscle, a connective tissue, an epithelial tissue, a wall of an abdominal passage, a liver, a kidney, a stomach, an appendix, an intestine, a pancreas, a lung, a heart, a bladder, a gall bladder, a brain, an endocrine gland, a reproductive gland, and a lymphatic gland.

3. The method of claim 1 wherein the genetic material is attached to microparticles.

4. The method of claim 3 wherein the microparticles are iron oxide particles ranging in size from 0.5 to 1 micron in diameter.

5. A method for delivering genetic material into a human or non-human subject, the method comprising the steps of:

contacting a tip of a microneedle with a solution comprising the genetic material;

penetrating the tip repeatedly into a site on a surface of a target tissue at an in situ site on the subject, to a depth in the tissue selected to allow contact between the tip and a plurality of target cells that can take up the genetic material, until a sufficient amount of the genetic material is delivered to the target cells such that a change in the subject attributable to the delivery of the genetic material is detectable.

6. A method as claimed in claim 5 wherein the target tissue is selected from the group consisting of skin cells and a wound.

7. A method as claimed in claim 5 wherein the target tissue is internal to the subject.

8. A method as claimed in claim 7 wherein the target tissue is selected from the group consisting of a smooth muscle, a striated muscle, a connective tissue, an epithelial tissue, a wall of an abdominal passage, a liver, a kidney, a stomach, an appendix, an intestine, a pancreas, a lung, a heart, a bladder, a gall bladder, a brain, an endocrine gland, a reproductive gland, and a lymphatic gland.

9. A method as claimed in claim 5 wherein less than about 500 micrograms of the genetic construct are delivered to the target cells.

10. A method as claimed in claim 5 wherein between about 20 and 200 micrograms of the genetic construct are delivered to the target cells.

11. A method as claimed in claim 5 wherein the penetrating step is repeated between about 500 and 5000 times per cm² of the surface of the target tissue.

12. A method as claimed in claim 5 wherein the penetrating step is repeated between 3000 and 4000 times per cm² of the surface of the target tissue.

13. A method as claimed in claim 5 wherein the tip penetrates the target tissue to a depth of less than 5 mm.

14. A method as claimed in claim 5 wherein the tip penetrates the target tissue to a depth of less than 3 mm.

15. A method as claimed in claim 5 wherein the genetic material encodes a non-native growth factor.

16. A method as claimed in claim 15 wherein the non-native growth factor is epidermal growth factor.

17. A method as claimed in claim 5 wherein the subject is non-human.

18. A method as claimed in claim 17 wherein the subject is selected from a group consisting of a dog, a cat, cattle, swine, a goat, and a sheep.

19. A method as claimed in claim 5 wherein the subject is a human.

20. A method as claimed in claim 5 further comprising the step of:

placing the microneedle in a concentrated solution of hydrochloric acid before the penetrating step.

21. A method for delivering genetic material into a target tissue of a subject, the method comprising the steps of:

contacting a tip of a microneedle with a solution comprising the genetic material;

penetrating the tip repeatedly into a site on a surface of the target tissue, to a depth selected to allow contact between the tip and a plurality of target cells that can take up the genetic material, until a sufficient amount of the genetic material is delivered to the target cells such that a change in the subject attributable to the delivery of the genetic material is detectable,

wherein the target tissue is selected from the group consisting of skin cells and a wound, the method further comprising the step of covering the target cells with a protective chamber.