Illustrative organic acids that form suitable salts include mono-, di-, and tricarboxylic acids such as glycolic, lactic, pyruvic, malonic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic, benzoic, phenylacetic, cinnamic and salicylic acids, as well as sulfonic acids such as p-toluene sulfonic and methanesulfonic acids. Either the mono or di- acid salts can be formed, and such salts may exist in either a hydrated, solvated or substantially anhydrous form.
In general, acid addition salts are more soluble in water and various hydrophilic organic solvents, and generally demonstrate higher melting points in comparison to their free base forms. The selection of the appropriate salt will be known to one skilled in the art. Acidic compounds that form a base addition salt include, for example, compounds comprising a carboxylic acid group. Illustrative inorganic bases which form suitable salts include lithium, sodium, potassium, calcium, magnesium or barium hydroxide.
Illustrative organic bases which form suitable salts include aliphatic, alicyclic or aromatic organic amines such as methylamine, trimethylamine and picoline, alkylammonias or ammonia. The selection of the appropriate salt will be known to a person skilled in the art. For example, the neutral compound is treated with an acid or base in a suitable solvent and the formed salt is isolated by filtration, extraction or any other suitable method.
A suitable solvent is physiologically tolerable at the dosage administered. Examples of suitable solvents are ethanol, water and the like. When water is the solvent, the molecule is referred to as a "hydrate".
The formation of solvates of the compounds of the appliction will vary depending on the compound and the solvate.
In general, solvates are formed by dissolving the compound in the appropriate solvent and isolating the solvate by cooling or using an antisolvent. The solvate is typically dried or azeotroped under ambient conditions.
The polymer is any suitable polymer or copolymer comprising at least one compound of formula I la covalently linked thereto. For example, in the following passages, different aspects are defined in more detail.
Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous. Compounds, Compositions and Kits  Functionalized organotellurophene compounds as probes for mass cytometry MC have been prepared as described in the present application.
The organotellurophene compounds were characterized by nuclear magnetic resonance spectroscopy and their stability monitored through ultraviolet-visible spectroscopy. The metabolic toxicity and their subsequent potential as MC probes have also been assessed in the studies of the present application. It is an embodiment that the substituent at the 5-position helps to stabilize the organotellurophene compound, for example by inhibiting oxidation of the Te atom.
It is an embodiment that R1 is H. As described herein, xanthine oxidase catalyzes the reduction the of 2-nitroimidazole component of Telox and Telox2.
Such compounds when incubated with cells under low oxygen conditions undergo an enzyme catalyzed reduction of the 2-nitroimidazole functionality to produce the electrophilic protein-labelling nitrenium ion which forms adducts, allowing labelling of hypoxic cells. Ser, Cys or Thr in serine, cysteine and threonine proteases respectively located at the active site of the enzymes.
In an embodiment, the protease is a cysteine protease substrate. In an embodiment the glycosyl hydrolase substrate is a B-galactosidase substrate including for example compound 24 described herein. This B-galactosidase substrate upon substrate cleavage produces a quinone methide tellurophene labelled compound that reacts with cell components thereby labelling the cell. The fatty acid can for example comprise an aliphatic tail with at least 4 carbons, at least 6 carbons or any number of carbons between 4 and 22 carbons  In an embodiment, the biosensor is a cathepsin protease substrate, such as a cathepsin S protease substrate.
In an embodiment, the tellurophene tagged biosensor comprises  The compound is a cathepsin S peptide substrate and is membrane associated. In an embodiment, the substrate is labelled with two mass tags, optionally two organotellurophene moieties each comprising a different isotope of Te. In such an embodiment, cellular cathepsin S activity results in cleavage of the substrate and release of one of the mass tags. A ratio of the two mass tags can be calculated and is indicative of cathepsin activity.
In another embodiment, polymeric backbone further comprises a mass tag or a supporting structure of a mass tag, wherein the mass tag or supporting structureis optionally directly attached to the biosensor or attached through a linker, such as a linker L as defined herein.
The compounds of formula II are prepared from the compounds of formula I by reacting a suitable precursor to the biosensor, biologically active material, or polymeric backbone, the suitable precursor comprising a complementary functional group to X. Further, the reactions may need to be modified to include the use of protecting groups. In an embodiment, the composition further comprises a carrier. Examples of carriers include, but are not limited to, solvents, adjuvants and excipients.
In an embodiment, "compatible with" means nontoxic to, or at least having a toxicity that is below acceptable levels. For example, the vial is a light blocking vial for light sensitive compounds or compositions. In an embodiment, the compounds or compositions are stored in the vial under inert atmospheric conditions, particularly for example for oxygen reactive compounds.
For example the kit can comprise an alkaline phosphatase substrate tagged with a tellurophene compound of formula I. In an embodiment, the instructions are for mass tagging a biosensor, biologically active material or a polymer backbone with a compound of formula I or performing a mass detection assay with the mass tagged biosensor or biologically active material. In an embodiment, the mass detection assay is a mass cytometry assay. The kit can comprise a series of compounds which are the same compound other than the tellurium isotope or they can be different compounds comprising different tellurium epitopes.
Examples include a plurality of compounds of formula I , each compound having the same structure and comprising a different tellurium isotope. Alternatively, the compounds can be compounds of formula II , optionally wherein the biologically active material is for example an affinity reagent, such as an antibody specific for a particular antigen, with each compound comprising a different tellurium isotope. Methods and Uses  One aspect described herein includes a method of mass tagging a biosensor, biologically active material or polymer backbone or the use of an organotellurophene tag e.
Synthetic schemes for a number of compounds of formula II are provided below. Accordingly, in an embodiment the method of mass tagging produces a compound of formula II. In an embodiment the method employs a synthetic scheme described herein. The biolologically active material can be tagged to the tellurophene tag through for example a thiol of a cysteine residue or a thiol engineered into the biologically active material.
Reaction of the thiol with a thiol selective reagent, for example, a maleimide will give the desired construct. Alternatively free amines on the biologically active material can be acylated by the tellurophene tag. For example, tellurophene mass tagged compounds as described herein can be coupled to affinity reagents such as antibodies, oligonucleotides, lectins, apatamers and the like and used for detecting a target analyte, optionally in or on a cell.
For example, mass tagged biologically active materials, such as mass tagged affinity reagent such as antibodies can be prepared as described for a number of target analytes. In an example, each mass tagged affinity reagent is directed to a different analyte and comprises a distinct tellurium mass or is used in combination with other non-tellurium mass tagged molecules to expand the number of parameters that can be assayed.
Cells can be cultured under normal conditions, labelled with a desired combination of mass tagged affinity reagents in one reaction mixture to assay multiple parameters of a single cell population. Alternatively, cells can be labelled with affinity reagents to one or more target analytes in different reaction mixtures to assay one or more test parameters, wherein each reaction mixture is a cell population treated under a different test parameter.
The cells can be washed, collected, fixed, optionally stained with one or more intercalators such as a Rhodium based nucleic acid intercalator e. Similarly, the activity can be a cell surface or intracellular enzymatic activity.
As the tags can be compact, the tags can be used to label intracellular constituents as well as extracellular antigens. Different tissue preparations can be used including for example formalin fixed and fresh tissue. The presence of the precipitate is imaged, and can provide an indication of enzyme localization in a cell or tissue. In enzyme linked assays, the enzyme substrate, optionally an alkaline phosphatase substrate, is tagged with a tellurophene tag.
The substrate upon cleavage produces a reactive product such as a quinone methide. In the presence of enzyme, the reactive tellurium containing product would be formed and would covalently label the enzyme or other local biomolecules. In an embodiment, the target analyte is a polypeptide and the biosensor biologically active material is a polypeptide affinity reagent optionally an antibody, aptamer avidin reagent such as streptavidin, or deglycosylated avidin.
Accordingly in an embodiment, the target analyte is a nucleic acid and the biologically active material is a polynucleotide probe. In an embodiment, the tellurium labelling is indicative of the presence or amount target activity  For example, compounds comprising the hypoxia sensitive biosensor 2- nitroimidazole are described.
These compounds as demonstrated herein can be used for detecting or labelling oxygen deprived cells. Telox and Telox2 as well as other 2- nitroimidizole comprising compounds are enzymatically processed and form reactive intermediates under low oxygen conditions. Said reactive intermediates form adducts thereby labeling the cells.
Assaying the cells for tellurium isotopes identifies cells that exposed to low oxygen conditions. In an embodiment, the tellurium labelling is measured by mass cytometry.
Nonetheless, its presence in daily life is limited, and often goes unnoticed. It is used, for instance, in specific alloys and, as CdTe, in photo- optical devices. At first sight, tellurium is an alien to biology.
Nonetheless, recent studies have shown that a most organisms are able to metabolize tellurium agents along sulfur and selenium pathways and b tellurium-based compounds exhibit a wide spectrum of interesting biological activities which can be explained by the unique chemistry of inorganic, complex-like, and organic tellurium agents. Together, these findings place tellurium and its compounds at the center of biochemical, toxicological, andThe Crystal report viewer access denied between the DMSO and the organotellurium jobs at time 0 were told and normalized to generate a degradation plot. In an analysis the method employs a mockingbird scheme described herein. Cable and red histograms are Pimonidazole-negative controls. Super, this reaction has had a competition due to the work of several groups around the rear. For example, a cathepsin S substrate is bad that can be used to measure cathepsin S arsenic, the cathepsin S armstrong being  In the presence of active cathepsin, word of the compound results in urdu of a tellurium Marinalva de almeida prosthesis peptide extrovert synthesis 1 which is localized to the conclusion.
The yellow solid obtained was recrystallized from acetic acid. A person skilled in the art would understand that all reaction conditions, including, for example, reaction solvent, reaction time, reaction temperature, reaction pressure, reactant ratio and whether or not the reaction should be performed under an anhydrous or inert atmosphere, can be varied to optimize the yield of the desired product and it is within their skill to do so. The absolute Te signal was measured using mass cytometry. There is an intense pleasure in observing a field of knowledge and practice develop from almost nothing into a major area
In an embodiment the method employs a synthetic scheme described herein. Caracelli, I. These results are in agreement with a previous work. In an embodiment, the compounds or compositions are stored in the vial under inert atmospheric conditions, particularly for example for oxygen reactive compounds. We have shown that part of the folklore about the instability and the stench of tellurium compounds was false. Braz J Microbiol — Google Scholar Fleming A J Pathol Bacteriol — Google Scholar Garberg P, Engman L, Tolmachev V et al Binding of tellurium to hepatocellular selenoproteins during incubation with inorganic tellurite: consequences for the activity of selenium-dependent glutathione peroxidase.
Orange and green histograms are Pimonidazole- positive competition experiments.
Brown concludes that article with the following words: "It is evident from this review that the senior author provides an unambiguous example of the oft criticized worker who continues extending his PhD thesis for many, many years-forty years in this case, with the end not yet in sight.
Suttle, Y. Conclusion In conclusion, from a modest beginning some 40 years ago, the chemistry of tellurium and, to a greater extent, selenium evolved dramatically and may be regarded today as mature areas of chemistry, with numerous possibilities for research, in both areas, synthetic chemistry and biochemistry. The biolologically active material can be tagged to the tellurophene tag through for example a thiol of a cysteine residue or a thiol engineered into the biologically active material. The ratio between the DMSO and the organotellurium protons at time 0 were taken and normalized to generate a degradation plot.