365-08-2

Basic information

• Product Name: thymidine 5'-(tetrahydrogen triphosphate)
• Synonyms: thymidine 5'-(tetrahydrogen triphosphate);[hydroxy-[hydroxy-[[3-hydroxy-5-(5-methyl-2,4-dioxo-pyrimidin-1-yl)-oxolan-2-yl]methoxy]phosphoryl]oxy-phosphoryl]oxyphosphonic acid;Thymidine-5''-triphosphoric acid;2'-Deoxythymidine triposphate;5-Methyl-dUTP;5'-TTP;Deoxy-TTP
• CAS NO: 365-08-2
• Molecular Formula: C₁₀H₁₇N₂O₁₄P₃
• Molecular Weight: 482.168263
• EINECS: 206-669-7
• Mol File: 365-08-2.mol

Chemical Properties

• Appearance: /
• Density: 1.922 g/cm³
• Storage Temp.: 2-8°C
• PKA: 0.80±0.50(Predicted)
• CAS DataBase Reference: thymidine 5'-(tetrahydrogen triphosphate)(CAS DataBase Reference)
• NIST Chemistry Reference: thymidine 5'-(tetrahydrogen triphosphate)(365-08-2)
• EPA Substance Registry System: thymidine 5'-(tetrahydrogen triphosphate)(365-08-2)

Safety Data

• Hazardous Substances Data: 365-08-2(Hazardous Substances Data)

Usage

Uses

Used in Molecular Biology Research:
Thymidine 5'-(tetrahydrogen triphosphate) is used as a substrate in various molecular biology techniques for DNA synthesis and amplification. It is essential for the in vitro replication of DNA, allowing researchers to study genetic material and perform experiments that require the synthesis of DNA.
Used in Diagnostics and Therapeutics:
In the medical field, Thymidine 5'-(tetrahydrogen triphosphate) is used as a diagnostic tool in molecular diagnostics, such as in the detection of specific DNA sequences or mutations. It is also employed in the development of therapeutic agents targeting DNA synthesis, which can be used to treat diseases caused by abnormal DNA replication or repair mechanisms.
Used in DNA Sequencing:
Thymidine 5'-(tetrahydrogen triphosphate) is used as a reagent in DNA sequencing techniques, such as Sanger sequencing, where it helps in the synthesis of DNA fragments that can be analyzed to determine the order of nucleotides in a DNA molecule.
Used in DNA Repair Mechanisms:
In cellular processes, Thymidine 5'-(tetrahydrogen triphosphate) plays a crucial role in DNA repair mechanisms, where it provides the energy and building blocks necessary for the accurate repair of damaged DNA, thus maintaining the integrity of genetic information.
Used in Genetic Engineering:
In the field of genetic engineering, Thymidine 5'-(tetrahydrogen triphosphate) is used as a component in various techniques that involve the manipulation of DNA, such as gene cloning, gene editing, and the construction of recombinant DNA molecules. It facilitates the incorporation of new genetic information into target organisms or cells, enabling the development of genetically modified organisms with desired traits.

Upstream product

• 63593-01-1 3'-O-benzyl-2'-deoxythymidine 
• 50-89-5 thymidine 
• 365-07-1 thymidine 5'-phosphate 
• 7791-71-1 5'-O-tritylthymidine 
• 1092560-23-0 C₁₅H₂₅N₂O₁₅P₃ 
• 1092560-22-9 C₁₄H₂₃N₂O₁₅P₃ 
• 90290-82-7 5'-triphosphate-3'-O-acetylthimidine 
• 1092560-21-8 C₁₃H₂₁N₂O₁₆P₃ 
• 1092560-20-7 C₁₈H₂₃N₂O₁₆P₃ 
• 93966-64-4 3'-benzoyl-5'-O-(4,4'-dimethoxytrityl)thymidine 
• 101527-40-6 5'-O-tert-butyldiphenylsilylthymidine 

Downstream Products

• 62012-79-7 dTDP-β-L-fucose 
• 38168-82-0 glyceric acid 1,3-biphosphate 
• 515138-66-6 3-O-methyl-dTDP-α-D-glucose 
• 515138-64-4 C₁₆H₂₅N₅O₁₅P₂ 
• 7132-58-3 dTDP-4-keto-6-deoxy-D-glucose 
• 16752-71-9 thymidine diphosphate 4-keto-6-deoxy-α-D-glucose 
• 86119-84-8 phosphoric acid 
• 2147-59-3 thymidine 5'-diphospho-β-L-rhamnose 
• 365-07-1 thymidine 5'-phosphate 
• 66-22-8 uracil 

Relevant articles and documents

PPN Pyrophosphate: A New Reagent for the Preparation of Nucleoside Triphosphates

Tris{bis(triphenylphosphoranylidene) ammonium} (PPN) pyrophosphate was accessed via aqueous precipitation and desiccation. The reagent was investigated as a replacement for highly hygroscopic alkylammonium salts in Ludwig-Yoshikawa reactions for the preparation of nucleoside-5′-triphosphates.

REVERSIBLE TERMINATORS FOR DNA SEQUENCING AND METHODS OF USING THE SAME

The present disclosure provides methods of sequencing polynucleotides and compounds, compositions for sequencing of polynucleotides, and synthesis of such compositions. The chemical compounds include nucleotides and their analogs which possess a sugar moiety comprising a cleavable chemical group capping the 3'-OH group and a base, but without covalently bounded dye. The cleavable chemical group is reactive to form covalent bond(s) with a dye used to confirm the presence of the expected base-pairing. The cleavable chemical group capping the 3'OH group can be removed together with the covalently bounded dye. Furthermore, after the cleavable chemical group is cleaved, the free 3'-OH group can be active in continued elongation. Example chemical compounds according to the present disclosure are shown as Formulas (II) and (V).

Method for synthesizing nucleotide or nucleotide analogue

The invention provides a method for synthesizing nucleotide or nucleotide analogue. The method includes combining nucleoside with phosphate. The pyrophosphate or tripolyphosphate undergoes nucleophilic substitution reaction under the catalysis of a phase transfer catalyst to obtain said nucleotide or nucleotide analogue, wherein said nucleoside has the structure represented by formula (I) or a stereoisomer, tautomer, pharmaceutically acceptable salt or prodrug thereof shown in formula (I).

MODIFIED NUCLEOTIDES AND USES THEREOF

Disclosed herein, inter alia, are compounds, modified nucleotides, compositions, and methods of using the same.

Lipophilic Triphosphate Prodrugs of Various Nucleoside Analogues

The antiviral efficacy of many nucleoside analogues is strongly dependent on their intracellular activation by host cellular kinases to yield ultimately the bioactive nucleoside analogue triphosphates (NTP). The metabolic conversion of nucleoside analogues into their triphosphates often proceeds insufficiently. We developed a nucleoside triphosphate (NTP) delivery system (the TriPPPro approach), in which the γ-phosphate is covalently modified by two different biodegradable masking units, one is the acyloxybenzyl (AB) moiety and the other is the alkoxycarbonyloxybenzyl (ACB) group. Such compounds formed NTPs with high selectivity by an enzyme-triggered mechanism in human T-lymphocyte CEM cell extracts loosing first the AB moiety, followed by the ACB group. This enables the bypass of all steps of the intracellular phosphorylation. This approach was applied here to convert some modestly active or even inactive nucleoside analogues into powerful biologically active metabolites. Potent antiviral activity profiles were obtained depending on the lipophilicity of the TriPPPro-NTP prodrugs against HIV-1 and HIV-2 replication in cultures of infected wild-type CD4+ CEM T-cells and more importantly in thymidine kinase-deficient CD4+ T-cells (CEM/TK-). This TriPPPro strategy offers high potential for future antiviral and antitumoral chemotherapies.

Synthetic method of nucleoside tetraphosphate

The invention discloses a synthetic method of nucleoside tetraphosphate. The synthetic method comprises the steps of carrying out selective phosphorylation reaction by virtue of nucleoside and a cyclic phosphorylation reagent, and carrying out oxidation and hydrolysis loop opening, so as to obtain nucleoside tetraphosphate. The structure of the cyclic phosphorylation reagent is represented by a formula I (shown in the description). According to the synthetic method, 5'-nucleoside tetraphosphate is selectively generated from nucleoside under the effect of the high-selectivity phosphorylation reagent, and 3'-OH (and 2'-OH) does not need to be protected in the process, namely that the generaiton of 3'(and 2'-)tetraphosphate can be effectively inhibited. Nucleoside tetraphosphate synthesized by virtue of the method has wide use ranges in the biology fields of DNA sequencing, labeling, extension and the like; currently, the selling prices is expensive, a synthetic method is complex, the reaction selectivity is poor; and the synthetic method provided by the invention is good in selectivity and easy in separation and purification, required experimental conditions are simple, and the synthetic processes are all conventional chemical reactions, so that the synthetic method is applicable to large-scale popularization and use.

COMPOSITIONS AND METHODS FOR SYNTHESIS OF PHOSPHORYLATED MOLECULES

The invention provides compositions and methods for synthesis of phosphorylated organic compounds, including nucleoside triphosphates.

P(V) Reagents for the Scalable Synthesis of Natural and Modified Nucleoside Triphosphates

Natural and modified nucleoside triphosphates impact nearly every major aspect of healthcare research from DNA sequencing to drug discovery. However, a scalable synthetic route to these molecules has long been hindered by the need for purification by high performance liquid chromatography (HPLC). Here, we describe a fundamentally different approach that uses a novel P(V) pyrene pyrophosphate reagent to generate derivatives that are purified by silica gel chromatography and converted to the desired compounds on scales vastly exceeding those achievable by HPLC. The power of this approach is demonstrated through the synthesis of a broad range of natural and unnatural nucleoside triphosphates (dNTPs and xNTPs) using protocols that are efficient, inexpensive, and operationally straightforward.

Chemical and enzymatic characterization of recombinant rabbit muscle pyruvate kinase

The stepwise synthesis of thymidine triphosphate (TTP) requires a kinase for phosphorylation in the last step. Because pyruvate kinase (PK) using phosphoenolpyruvate (PEP) as substrate can regenerate adenosine triphosphate and phosphorylate thymidine diphosphate as well, we chose this enzyme for the synthesis of TTP via an enzymatic cascade reaction. The metalloenzyme PK shows pronounced promiscuity and therefore fits well to the conditions of this reaction. PK commonly used today is isolated from rabbit muscle. We cloned and expressed the respective open reading frame in Escherichia coli, purified, and characterized the His-tagged recombinant enzyme. The enzyme has an activity optimum at 37 °C and in the pH range from 7.4 to 7.8. Km constants conformed well with the isolated native enzyme for adenosine diphosphate (ADP) to 0.37±0.02 mm and for PEP to 0.07±0.01 mm. The recombinant enzyme shows the following range in its substrate specificity: ADP>dADP>dGDP>dCDP>thymidine diphosphate (TDP). It allows the phosphorylation of TDP to TTP in high yield (up to 95 % ). The metal ions Mg2+ and K+ are necessary for full enzymatic activity. The addition of transition metal ions such as Mn2+, Cu2+, Co2+, and Ni2+ reduces activity. Storage of the enzyme at -20 °C retains full activity.

An improved protection-free one-pot chemical synthesis of 2′-deoxynucleoside-5′-triphosphates

□ A facile, straightforward, reliable, and an efficient method for the gram-scale chemical synthesis of both purine deoxynucleotides such as 2 ′-deoxyguanosine-5 ′-triphosphate (dGTP) and 2 ′- deoxyadenosine-5′-triphosphate (dATP) and pyrimidine deoxynucleotides such as 2 ′-deoxycytidine- 5 ′-triphosphate (dCTP), thymidine-5 ′-triphosphate (TTP), and 2 ′-deoxyuridine-5 ′-triphosphate (dUTP) starting from the corresponding nucleoside is described. This improved "one-pot, three step"Ludwig synthetic strategy involves the monophosphorylation of nucleoside followed by reaction with tributylammonium pyrophosphate and hydrolysis of the resulting cyclic intermediate to provide the corresponding dNTP in good yields (65%-70%). Copyright Taylor and Francis Group, LLC.