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[Federal Register Volume 70, Number 25 (Tuesday, February 8, 2005)]
[Notices]
[Pages 6702-6704]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 05-2364]

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DEPARTMENT OF HEALTH AND HUMAN SERVICES

National Institutes of Health

Government-Owned Inventions; Availability for Licensing

AGENCY: National Institutes of Health, Public Health Service, DHHS.

ACTION: Notice.

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SUMMARY: The inventions listed below are owned by an agency of the U.S. 
Government and are available for licensing in the U.S. in accordance 
with 35 U.S.C. 207 to achieve expeditious commercialization of results 
of federally-funded research and development. Foreign patent 
applications are filed on selected inventions to extend market coverage 
for companies and may also be available for licensing.

ADDRESSES: Licensing information and copies of the U.S. patent 
applications listed below may be obtained by writing to the indicated 
licensing contact at the Office of Technology Transfer, National 
Institutes of Health, 6011 Executive Boulevard, Suite 325, Rockville, 
Maryland 20852-3804; telephone: 301/496-7057; fax: 301/402-0220. A 
signed Confidential Disclosure Agreement will be required to receive 
copies of the patent applications.

Methods for Treating Active Uveitis

Robert Nussenblatt (NEI) and Thomas Waldmann (NCI), Zhuqing Li (NEI), 
Ronald Buggage (NEI).
U.S. Provisional Patent Application No. 60/616,760 filed 06 Oct 2004 
(DHHS Reference No. E-328-2004/0-US-01).
Licensing Contact: Susan Carson; 301/435-5020; [email protected].

    Intraocular inflammatory disease (uveitis) is characterized by pain 
and a decrease in vision that can lead to blindness if not treated 
appropriately. The incidence and prevalence of the disease are 
approximately 52/100,000 and 112/100,000, and this translates into an 
incidence of 151,000 per year and a prevalence of 322,000. The numbers 
are expected to increase as the population ages. Treatment of severe 
uveitis often focuses on the control of the inflammatory symptoms using 
high dose corticosteroids, cytotoxic drugs or other immunosuppressive 
agents and there is a need for therapies that reduce the major side 
effects associated with the prolonged use of systemic steroids (e.g. 
hyperglycemia, osteoporosis and loss of immunocompetence).
    Daclizumab is a humanized anti-Tac (HAT) antibody that specifically 
binds to the alpha subunit (CD25 or Tac subunit) of the human high 
affinity interleukin-2 (IL-2) receptor expressed on the surface of 
activated lymphocytes. Dr. Nussenblatt and colleagues at the NEI have 
previously shown that daclizumab can be used to successfully treat 
quiescent uveitis. Long term daclizumab therapy at a dose of 1mg/kg can 
be used instead of standard immunosuppressive agents to treat severe 
uveitis for more than 4 years with no adverse effects attributable to 
the medication, and subcutaneously administered daclizumab also 
appeared to be clinically effective. However, subjects with active 
uveitis were less likely under this regimen to have their disease 
controlled (J. Autoimmunity (2003) 21, 283-293).
    The present invention targets patients with refractory, active 
uveitis and consists of a high dose intravenous induction therapy using 
daclizumab at two different doses and times followed by a longer term 
maintenance therapy. Positive therapeutic effects have been seen with 
this protocol in a small group of patients within 4-6 weeks after the 
initiation of therapy. As previous work indicated that IL-2R receptors 
have a slow turnover rate on CD4 positive subpopulation of lymphocytes, 
a possible mechanism of action of this new protocol is saturation of 
CD25 (TAC) receptors on cells in sequestered sites.
    Available for licensing are methods directed to this treatment of 
active uveitis using a high dose pulsatile induction protocol of an 
interleukin-2 (Il-2) receptor antagonist. Methods are also provided for 
the treatment of corneal transplant rejection, limbal stem cell 
rejection following transplantation, optic neuritis and dry eye.

Novel Thermostable Y-Family DNA Polymerases

Roger Woodgate (NICHD), John P. McDonald (NICHD), and Wei Yang (NIDDK).
U.S. Provisional Patent Application No. 60/573,684 filed 20 May 2004 
(DHHS Ref No. E-166-2004/0-US-01); U.S. Provisional Patent Application 
No. 60/623, 490 filed 29 Oct 2004 (DHHS Ref No. E-166-2004/1-US-01).

[[Page 6703]]

Licensing Contact: Susan Carson; 301/435-5020; [email protected].

    Y-family polymerases are able to bypass lesions in DNA that would 
otherwise block replication by high fidelity DNA polymerases and are 
key to the effective study of ancient DNA and for use in forensic 
medicine. These enzymes are ubiquitous and are found in all kingdoms of 
life: bacteria, archaea and eukaryotes. The number of proteins related 
to the Y-family polymerases is well over 200 orthologs and despite 
being closely related at the phylogenetic level, the few polymerases 
now characterized, each show a unique set of properties including 
processivity, fidelity, and the ability to bypass certain types of DNA. 
Y-family polymerases from thermostable organisms are of particular 
interest because the enzymes isolated from such species tend to be more 
stable, easy to work with and may have more utility in assays at higher 
temperatures, such as Polymerase Chain Reaction (PCR). For example, the 
thermostable archeal Sulfolobus solfataricus DinB-like polymerase Dpo4 
can bypass lesions by generally inserting the correct complementary 
nucleotide opposite a variety of damaged bases and can, under 
appropriate conditions substitute for Taq polymerase in PCR 
applications [NAR (2001) 29, 4607-4616; DHHS Ref. No. E-232-2001/0]. 
Additionally, functional and structural organization of this family of 
polymerases permits domain swapping designed to optimize specific 
properties of use in novel applications [J. Biol. Chem. (2004) 279, 
32932-32940].
    Dr. Woodgate's group at the National Institute of Child Health and 
Development have expanded their earlier work and have now discovered 
several additional thermostable dpo4 homologs from other strains found 
in the Sulfolobaceae family, some of which have optimal growth 
temperatures higher than 80[deg]C. These novel DinB-like proteins have 
thermostable DNA polymerase activity and are capable of: (1) PCR 
amplifications over 1kb in length, (2) replication past DNA lesions 
such as abasic sites and CPD (cis-syn cyclobutane pyrimidine dimer) 
lesions and (3) incorporation of several different labeled DNA 
nucleotides into DNA during replication. These enzymes may therefore be 
a good substitute for Taq polymerase in applications utilizing 
fluorescent nucleoside triphosphate derivatives. These lesion-bypassing 
Dpo4-like polymerases could also be included along with a conventional 
thermostable polymerase in a PCR protocol designed to amplify old or 
damaged DNA samples which could greatly increase recoverability, 
accuracy and length of products. Other applications could include 
labelling or tagging DNA, real-time PCR, detection of SNPs, mismatches 
or DNA lesions, mutagenic PCR, directed-evolution methods and expanding 
the ``DNA alphabet'' utilizing non-natural nucleotides.
    Available for licensing are seven novel Y-family polymerases. 
Claims are directed to these sequences and chimeras, as well as to 
methods of identifying other Y-family polymerases and generating other 
chimeric Y-family polymerases and methods of use. These enzymes and 
methods of identifying and generating novel Y-family polymerases should 
be of interest to forensic DNA service companies as well as to research 
reagent companies pursuing novel thermophlic enzymes for use in ancient 
and damaged DNA analysis and for novel applications with modified 
nucleotides.
    In addition to licensing, the technology is available for further 
development through collaborative research with the inventors via a 
Cooperative Research and Development Agreement (CRADA).
    Related technologies available for licensing as research tools 
include: DHHS Ref. No. E-232-2001/0 (dpo4 Y-family polymerase) and DHHS 
E-229-2001/0 (pol iota Y-family polymerase).

Rapid, Efficient In Vivo Site-Directed Mutagenesis Using 
Oligonucleotides

Francesca Storici, Michael A. Resnick, Lysle Kevin Lewis (NIEHS).
PCT Application No. PCT/US02/23634 filed 26 July 2002, which published 
as International Publication No. WO03/012036 on 13 Feb 2003 (DHHS 
Reference No. E-204-2001/0-PCT-02).
National Stage Entry: EPC, CA, AU, US, JP.
U.S. Patent Application No. 10/484,989 filed 26 Jan 2004 (DHHS 
Reference No. E-204-2001/0-US-07).
Licensing Contact: Susan Carson; 301/435-5020; [email protected].

    The rapid modification of genes provides opportunities to study 
gene function and evaluate drug responsiveness. Scientists at the 
National Institute for Environmental and Health Sciences have developed 
a new system in yeast, delitto perfetto, which provides for rapid, 
efficient and accurate in vivo genomic mutagenesis using 
oligonucleotides (IROs) and involves the complete removal of the 
heterologous sequence previously integrated at the target locus (Nature 
Biotechnol. (2001) 19, 773-776). They have demonstrated that synthetic 
oligonucleotides can target a desired mutation to almost any 
chromosomal locus where a marker cassette has been previously 
integrated. The oligonucleotides, which are designed with short 
sequence homology to sites up- and down-stream of the marker cassette, 
replace the marker cassette with the chosen mutation without leaving 
any heterologous sequence in the targeted locus. Since the system 
always provides selection for the clones containing the desired 
mutation, it can be used to generate any kind of modification: i.e., it 
is not constrained by the generation of mutations that provide a 
detectable phenotype. Additionally, induction of double-strand breaks 
(delitto perfetto-DSB) in vivo before standard transformation 
procedures provides 1,000 to 10,000 fold stimulation of oligonucleotide 
targeting, resulting in 5-20% of all cells in the population being 
efficiently targeted by small oligonucleotides (PNAS (2003), 100, 
14994-14999). With such a high stimulation of targeting even gross 
rearrangements, like large DNA deletions, chromosome fusions, 
circularizations, reciprocal or non reciprocal translocations are 
obtained with high frequency and direct selection.
    The core invention is a novel self-cloning system for simple and 
high-throughput in vivo site-directed mutagenesis applicable to all 
organisms capable of homologous recombination and developed in the non-
pathogenic yeast Saccharomyces cerevisiae. Since changes are created 
through a self-cloning process, this system could represent a highly 
versatile tool to generate modifications of genes in yeasts for 
commercial application in the food and beverage industries (such as, 
baking, brewing, wine and sake) without the resulting organisms being 
classified as GMO (genetically modified organisms). This approach could 
also be well positioned within drug discovery protocols where the need 
to mutagenize particular target sequences forms an integral part of the 
drug development process.
    Delitto perfetto-DSB is efficient for targeting homologous 
sequences that are close or distant to the DSB and in the presence of a 
competing homologous chromosome in diploid cells, and can strongly 
stimulate recombination with single-stranded DNA, without strand bias. 
The mechanism of DSB repair with oligonucleotides follows primarily a 
single-strand annealing pathway of recombinational repair. This novel 
system is also independent of restriction sites, requires minimal 
sequence

[[Page 6704]]

analysis. This method has been used in S. cerevisiae for many yeast 
chromosomal genes and the human gene p53 and has obvious potential for 
use with YAC and TAR clones. Claims are directed to several methods for 
generating DNA nucleic acid mutations in vivo and are applicable to any 
organism that has a homologous recombination system, as well as to 
kits. This methodology is available for licensing and is a highly 
versatile tool of direct use to drug discovery, pharma and research 
reagent companies as well as to companies working with industrial yeast 
strains.
    In addition to licensing, the technology is available for further 
development through collaborative research with the inventors via a 
Cooperative Research and Development Agreement (CRADA).
    Related technologies also available for licensing include: DHHS 
Ref. No. E-121-1996/0-US-06, Transformation-Associated Recombination 
Cloning (U.S. Patent No. 6,391,642 issued 21 May 2002); and DHHS Ref. 
No. E-262-1984/0-US-03, Process for Site Specific Mutagenesis Without 
Phenotypic Selection (U.S. Patent No. 4,873,192 issued 10 Oct 1989).

The Whey Acidic Protein (WAP) Promoter and Its Use to Express 
Therapeutic Proteins in the Milk of Transgenic Mammals

Lothar Hennighausen (NIDDK), Heiner Westphal (NICHD), et al. U.S. 
Patent No. 6,727,405 issued 27 Apr 2004 (DHHS Reference No. E-411-1987/
0-US-03).
Licensing Contact: Susan Carson; 301/435-5020; [email protected].

    Transgenic animals can be engineered to express complex human 
proteins at high concentrations in milk. Protein replacement therapy is 
often the only treatment available for congenital diseases such as 
hemophilia or lysosomal storage disease, and the cost of treatment can 
be high with the therapeutic protein market estimated to reach more 
than $50 billion by 2010.
    U.S. Patent No. 6,727,405 has recently been issued (expiry date 
2021) to NIH scientists and their collaborators. This patent provides 
for a non-human mammal such as mouse, sheep, pig, goat and cow whose 
genome contains a DNA sequence comprising a milk serum protein (whey 
acidic protein) promoter linked to a heterologous gene sequence and 
secretory peptide, as well as methods for producing a secreted protein 
into the transgenic animal's milk and claims directed to the DNA 
construct. The invention permits the production of any desired protein 
in an easily maintained, stable, mammalian bioreactor, which is capable 
not only of producing the desired protein in milk, but can also pass 
the ability to do so to its female offspring. Although other methods of 
obtaining recombinant protein products are available, these require 
inefficient, expensive purification of the protein from the blood or 
from cell culture media and there remains a need for an efficient and 
cost effective method for producing therapeutic proteins.
    This WAP promoter platform technology provides a viable alternative 
to other milk protein promoters and is available for non-exclusive 
licensing.

    Dated: January 31, 2005.
Steven M. Ferguson,
Director, Division of Technology Development and Transfer, Office of 
Technology Transfer, National Institutes of Health.
[FR Doc. 05-2364 Filed 2-7-05; 8:45 am]
BILLING CODE 4140-01-P
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