Invented by Chad A. Mirkin, Amy S. Paller, David A. Giljohann, Northwestern University
The Northwestern University invention works as followsThe present invention relates to compositions and methods for delivering an oligonucleotide-functionalized nanoparticle.
Background for Delivery of oligonucleotide-functionalized nanoparticles
The introduction of genetic material to cells and tissues, for the control of gene expression, has had a significant impact on research in gene pathways and functions and offers promise for therapeutic applications. Genetic level approaches have inherent specificity that is not possible with most drugs. Clinical trials are underway for siRNAs, which hold great promise in the future as therapeutic tools. They target a variety of clinical issues including cancer. Gene silencing can be more cost-effective and lead to a down-regulation in protein function and expression. It is also more specific than small-molecule inhibitors. Small molecule therapy, on the other hand, does not distinguish between mutant or normal gene products. Each melanoma has a unique genetic signature, which can be determined by identifying hotspot mutations in the melanoma, through direct gene sequencing or gene amplification assays. The siRNA is taken up by a wide range of cells. However, targeted gene therapy only affects cells that have a mutated or activated gene.
Oral delivery of siRNAs faces the same challenges as oral delivery of proteins. The degradation of nucleic acid and low bioavailability in the gastrointestinal tract is a major obstacle. Conventional siRNAs are rapidly degraded and do not reach their targets, even when delivered intravenously. Topical nucleic acid application offers therapeutic benefits, both in suppressing genes within lesional skin (for instance and without limitation to treat skin metastases) and in transdermal delivery of internal targets. The application is easy to control and painless, and the skin is easily accessible. The epidermis’s effective physical barrier is mainly located in the outermost epidermis layer, stratum corneum. It also extends to the deeper epidermis. This epidermal layer protects from water loss, both inside and outside. It also blocks the entry of other substances such as nucleic acid. The use of mechanical approaches such as ultrasounds, lasers and injections to penetrate the mouse stratum Corneum and deliver siRNA into the skin is possible, but requires specialized equipment and can potentially damage the skin. These challenges highlight the need for a transdermal system that can be easily applied to deliver suppressive nucleic acid.
Direct targeting of a skin condition is the ideal model for gene suppression therapy. The commercially available materials for suppressing genes in vitro are only marginally effective at delivering genetic material to primary cultured cell types, such as melanocytes and keratinocytes. The outer layers of the skin also function as an anatomical barrier, which traditionally prevents nucleic acid and protein penetration into the skin, and from the dermis into the circulation. [Prausnitz, et. al., Nat Biotechnol, 26: 1261-1268, (2008)]. This layer is difficult to penetrate in order to transfer enough oligonucleotides.
The skin is the largest body organ and has three layers, the epidermis dermis and subcutaneous tissue. The epidermis, or outer layer of the skin, is the most important part. Different types of skin have different thicknesses of epidermis. The epidermis is thinnest at the eyelids (0.05 mm) and thickest at the palms and soles (1,5 mm). The epidermis is made up of 4 layers, each with progressively more distinct cells. The layers are named from bottom to top:
The cells in the bottom layer (the stratum basale) are shaped as columns. This layer is where cells divide, pushing already formed cells up to higher layers. As cells progress into higher layers they flatten out, mature, and finally?die? The cells are shed when they flatten, mature and eventually?die? The stratum Corneum is the top layer of epidermis. It is composed of skin cells that have been flattened and shed.
The present disclosure provides compositions and methods for delivering an oligonucleotide-functionalized nanoparticle.
In some embodiments, the present disclosure provides a dermal composition comprising an oligonucleotide-functionalized nanoparticle (ON-NP) and a dermal vehicle.
Also provided by the present disclosure is a method of dermal delivery of an oligonucleotide-functionalized nanoparticle comprising the step of administering a composition comprising the oligonucleotide-functionalized nanoparticle and a dermal vehicle to the skin of a patient in need thereof.
In one aspect, the delivery of the oligonucleotide-functionalized nanoparticle is transdermal. In another aspect, the delivery of the oligonucleotide-functionalized nanoparticle is topical. In another aspect, the delivery of the oligonucleotide-functionalized nanoparticle is to the epidermis and dermis after topical application.
In some embodiments, a dermal vehicle includes an ointment. Aquaphor is an ointment in some aspects.
In another embodiment, a method of regulating gene expression is provided comprising the step of administering a therapeutically effective amount of a composition comprising an oligonucleotide-functionalized nanoparticle to skin under conditions wherein the oligonucleotide-functionalized nanoparticle hybridizes to a target and regulates gene expression.
In some aspects, a polynucleotide is the target. In some aspects, RNA is the polynucleotide. The target can be a polypeptide in some cases.
In other embodiments, administration of the composition alleviates a skin condition.
In various embodiments, skin disorders are selected from a group that includes cancer, genetic disorders, aging and infection.
The cancer can be selected in some cases from the group of basal cell cancer, squamous-cell carcinoma, breast cancer and melanoma.
In yet further embodiments, the targeted is a product of a gene expressed by one selected from a group consisting primarily of Ras, B-Raf and I?B?.
In other aspects, the target is a gene product that comprises a mutation, said gene product being expressed by a gene selected from a group consisting K5, K14, K1, m-Tor, H-Ras, and m-Tor. The target can also be a genetic product that contains a mutation. This gene product is expressed by one of the genes selected from K5, K14 K1, K10 H-Ras or m-Tor.
In some embodiments the aging disorder selected is from the group of UV-damage or progeria. In some aspects, the gene product is expressed by a selected gene from the group of matrix metalloproteinase-1 or progerin.
In some cases, the inflammation may be due to psoriasis. Interleukin 23 is the target in some cases.
In one embodiment, viral infection causes warts. “In some aspects, E6/E7 is the target.
In other embodiments, cosmetic disfigurement may be selected from a group including seborrheic lesions, epidermal nevi, and pigmented lesions. The target can be a genetic product containing a mutation. This gene product is expressed by one of the genes FGFR3, B-Raf, K10, and the like.Click here to view the patent on Google Patents.