Anticancer, Inc.

The United States of America, As Represented by the Secretary, Department of Health and Human Services

Yuying Tan

Mingxu Xu

Qinghong Han

Shukuan Li

Shigeo Yagi

Li Tang

Takashi Chishima

Xinghua Sun

Zhijian Yang

Valeryi Lishko

Martin Lenz

Kenji Miki

Ping Jiang

Huiyan Guo

Robert M Hoffman

Andrew W Perry

Quinghong Han

Marcin Lenz

Sant Chawla

Levy Kopelovich

Kei Kawaguchi

A composition and method for lowing serum and plasma levels of methionine by oral administration. The composition includes a recombinant methioninase enzyme and a cofactor (pyridoxal-L-phosphate). Methods of use describe methods for treatment of cancer, including malignant melanoma, by oral administration of the methioninase composition. Methods for chronic suppressive therapy of melanoma and other cancers are described. Because reduction of plasma methionine levels is effective in treating other conditions, including diabetes and conditions associated with aging, the use of the methods described herein includes treatment of these and other conditions.

Orally administered composition to lower serum methionine levels and method of use

A composition, method and kit for performing a two-reagent enzymatic homocysteine assay, wherein a single homocysteinase enzyme and a Schiff-based conjugate of N,N-dibutyl-p-phenyldiamine (DBPDA) with pyridoxal 5'-phosphate (PLP) are used to measure total homocysteine in plasma or serum. The assay measures a chromophore reaction product of HS and the DBPDA released from the Schiff-base conjugate in the presence of a Fecontaining compound. The resulting chromophore may be measured absorbance or fluorescence spectrophotometry.

Schiff-base conjugate of N, N-dibutyl-P-phenylenediamine with pyridoxal 5'-phosphate for improved homocysteine assays using pyridoxal 5&#x2032;-phosphate-dependent enzymes

The present invention relates to highly conjugated proteins and methods for making such proteins. In particular, the present invention relates to methods for linking additional sites to a protein for conjugation with activated polyethylene glycol (PEG) linkers, without denaturing the protein. The invention also relates to highly conjugated proteins with decreased immunogenicity and increased circulating half-life.

Methods for increasing protein polyethylene glycol (PEG) conjugation

A method is described to identify secreted proteins identified with stages of malignancy of cancer. The proteins are initially identified by trapping them with a fluorescent protein containing vector that can insert in any gene. The secreted proteins are initially identified by their fluorescence. Secreted proteins identifying tumors with specific degrees of malignancy are isolated to determine if they can serve as markers of cancer progression.

Methods for identifying markers for early-stage human cancer, cancer progression and recurrence

Infection bycan be treated by administering methioninase optionally in combination with additional therapeutic agents, such as antibiotics.

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Methods of obtaining faithful expression libraries from tissue samples comprise extraction of RNA from intact tissue cultured in three-dimensional sponge-gel based histocultures.

Expression profiling based on histocultures

A method to determine a total cysteine in biological fluids utilizes similarly treated portions of the fluid with a homocysteinase and a non-specific desulfurase.

Total cysteine assay

Methods for inhibiting the growth of tumor cells by combination treatment with a selenium-containing prodrug and an enzyme for which it is a substrate are described. The treatment also enhances the effect of anti-tumor agents.

Selenium-containing pro-drugs for cancer therapy

A method to follow the progression of metastasis of a primary tumor, which method comprises removing fresh organ tissues from a vertebrate subject which has been modified to contain tumor cells that express GFP and observing the excised tissues for the presence of fluorescence is disclosed. The fluorescence can also be monitored by observing the tissues in situ. Vertebrate subjects which contain GFP producing tumors are useful models to study the mechanism of metastasis, as well as to evaluate candidate protocols and drugs. In addition, subjects already harboring tumors can be treated so as to modify the endogenous tumors to contain GFP. This permits clinical applications. Finally, by injecting a contrast dye into a subject harboring a GFP-labeled tumor, angiogenesis in the tumor can be observed directly.

Metastasis models using green fluorescent protein (GFP) as a marker

A method to detect and measure the concentration of hydrogen peroxide comprises adding reagents consisting essentially of peroxidase and a dialkyl phenylenediamine and detecting any color generated by the interaction of hydrogen peroxide with these reagents.

Method to determine hydrogen peroxide

Compositions and methods which ameliorate the virulence of bacterial infection are described wherein the active ingredient modulates transmethylation reactions in bacterial cells. Particularly useful compounds are inhibitors of S-adenosyl methionine synthetase (SAMS), of S-adenosyl homocysteine hydrolase (SAHH) and of transmethylases.

Modulators of methylation for control of bacterial virulence

A depletion method to inhibit tumor growth includes introducing a viral expression system capable of expressing methioninase or a fusion protein containing methioninase into a tumor contained in a vertebrate subject or cells thereof. The fusion protein may contain a fluorescent protein to permit monitoring of the completeness of the depletion method. The fusion protein can be used in vivo as well as in vitro screening protocols which employ the viral expression system. The expression system includes control sequences and means to integrate the nucleotide sequence into the genome of a host cell for expression. The method may also include treating the cells with isolated methioninase and/or with a therapeutic cell. The depletion method can be with other known therapies, such as maintaining the animal having the tumor on a methionine depleted diet.

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San Diego, CA, United States of America

Yuying Tan