Therapeutics

This invention allows the engineering of semi-synthetic lantibiotics to a much larger extent than using typically practiced in vivo methods. The method is the first in vitro reconstruction lantibiotic photosynthesis and introduces non-proteinogenic amino acids.

Lantibiotics are peptide-based compounds produced by bacteria that live on lactic acid. They are used as natural antibiotic food preservatives as an alternative to chemical reagents and have been used for half a century without any significant signs of lantibiotic-resistant bacteria popping up.

Researchers at the University of Illinois have isolated and purified the LctM enzyme, responsible for synthesis of lacticin 481. This approach, the first in vitro reconstruction of lantibiotic biosynthesis, allows for lantibiotic engineering and opens the door to possible generations of new lantibiotics with greatly increased yields and improved activities.

Applications 

• Food preservation
• Industrial enzyme production
• Bioengineering of novel lantibiotics

Benefits

• Greater Production Flexibility: In vitro methodology allows for lacticin 481 production using semi-synthetic substrates.
• Avoids Complications of In Vivo Production: In vitro methods avoid product cytotoxicity, degradation and bypasses many of the In vivo regualtory complications.
• Better Resistance Profile: Lantibiotics have been used for half a century in more than 40 countries without any significant signs of lantibiotic-resistant bacteria appearing.
• More Possibilities: The method works well with combinatorial techniques, increasing the number of substrates that can be used in production of lacticin 481. In addition, isolation of the LctM enzyme allows for more efficient synthesis with greater structural and functional tolerance of the biosynthetic pathway. 

Dr. Zimmerman's lab has developed a series of bioactive compounds for the treatment of myotonic dystrophy. The compounds disrupt the MBNL-(CUG) foci in vivo and partially rescue the missplicing of cTNT and IR mRNAs. These novel compounds, utilizing bisamidinium-based inhibitors, show a 1000-fold greater inhibition potency than a previous compound and are able to degrade the causative agent of the disease.

Malignant melanoma is a cancer that is refractory to current chemotherapeutic treatments. The average survival time for patients is 6 months. This coupled with the alarming rise in incidence of melanoma (46,170 cases of in situ melanoma are expected to be newly diagnosed in 2005) warrants research into novel means to treat melanoma. Through the synthesis of a combinatorial library of potential proapoptotic agents, a novel class of small molecules (triphenylmethylamides, TPMAs) have been identified that are effective agents in killing melanoma cells, as well as other cancer cells. The majority of anticancer drugs operate through induction of cell cycle arrest and cell death in either the DNA synthesis (S) or mitosis (M) phase of the cell cycle. In contrast, these TPMAs, act in the G1 phase of the cell cycle. Many of the TPMAs are quite potent inducers of apoptotic death in melanoma cell lines (IC50 of 0.5 ?M), and importantly, some TPMAs are comparatively nontoxic to normal cells isolated from the bone marrow of healthy donors. Furthermore, the TPMAs dramatically reduce the level of active nuclear factor ?-B (NF?B) in the cell; NF?B is known to be constitutively active in melanoma, and this activity is critical for the proliferation of melanoma cells and their evasion of apoptosis. With regard to status of development, multiple generations of compounds have been tested and have shown efficacy with primary melanocytes, while these same compounds have shown to be largely non-toxic in initial rodent studies assessing changes in body weight and death. In addition, the National Cancer Institute?s 60-cell line screen has demonstrated activity of the compounds against a broad range of cancer types. Additional detailed information about this technology can be found in the recently published article: Synthesis and Identification of Small Molecules that Potently Induce Apoptosis in Melanoma Cells through G1 Cell Cycle Arrest (JACS 2005, 127, 8686-8696)

Applications

• Chemotherapeutic treatment of Melanoma Chemotherapeutic treatment against other cancers, such as breast, colon, non-small cell lung carcinoma (NSCLC), etc.
• As a research reagent that arrests cells in G1. 

Benefits

• Potent death inducers in melanoma cell lines with potencies in the 0.5 - 1.0 M range
• Markedly less (8-12-fold) toxicity to the non-cancerous human bone marrow cells
• National Cancer Institute's 60-cell line screen demonstrated the compounds are quite active against a broad spectrum of cancers and confirmed the IC50 value of 600 nM against the UACC-62 melanoma cell line Initial animal testing suggests that mice can tolerate concentrations as high as 200 mg/kg Efficacy and water solubility are being increased as additional chemical modifications are made with later generation compounds 

New anti-infectives are required now more than ever as resistance to existing drugs increases in prevalence. Enzymes unique to bacteria or parasites are potential drug targets with minimal side effects because they are not present in humans. One such source of potential drug targets is the isoprenoid biosynthesis pathway. Many pathogenic bacteria and malaria parasites use isoprenoids in their cell walls, to protect against the human immune system, and for other functions. This pathway is not present in humans and thus is an excellent target for new anti-infective drugs. Scientists from the University of Illinois at Urbana-Champaigns Departments of Chemistry and Biophysics have identified a class of novel chemical entities that are capable of inhibiting two key enzymes, GcpE and LytB, in the isoprenoid biosynthesis pathway. These compounds are able to inhibit the isoprenoid biosynthetic pathway at concentrations far lower than any other known inhibitors and have the potential to treat a wide-range of infectious disease caused by both bacteria and malarial parasites. In addition, it may be possible to use these compounds for the treatment of cancers, via immune system activation.

Details

This invention includes a class of novel chemical entities composed of similar geometries and bonds. They inhibit through a unique organometallicinteraction that has not been previously described.

Applications

Currently, the compounds are being evaluated for their ability to act as:

• Broad spectrum anti-infectives
• Cytotoxic cancer drugs
• Other compounds produced using similar chemistry may have a wide range of medical applications

Benefits

• Novel Mechanism: These compounds act on the isoprenoid pathway using a chemical interaction that has not been previously described.

Continued work on these compounds is designed to identify new applications of this interaction, beyond anti-infectives.