21679-14-1

Basic information

• Product Name: Fludarabine
• Synonyms: 2-Fluoroadeninearabinoside;9-beta-d-arabinofuranosyl-2-fluoro-9h-purin-6-amin;9-beta-d-arabinofuranosyl-2-fluoro-adenin;f-ara-a;6-AMINO-9 BETA-D-ARABINOFURANOSYLFLUOROPURINE;9-bata-d-arabinofuranosyl-2-fluoroadenine;9-BETA-D-ARABINOFURANOSYL-2-FLUORO-9H-PURIN-6-AMINE;9-BETA-D-ARABINOFURANOSYL-2-FLUOROADENINE
• CAS NO: 21679-14-1
• Molecular Formula: C₁₀H₁₂FN₅O₄
• Molecular Weight: 285.23
• EINECS: 244-525-5
• Product Categories: Intermediates & Fine Chemicals;Pharmaceuticals;API;F-ara-A, NSC 118218;Anti-cancer&immunity;Inhibitors
• Mol File: 21679-14-1.mol

Chemical Properties

• Melting Point: 265-268°C
• Boiling Point: 747.3 °C at 760 mmHg
• Flash Point: 405.8 °C
• Appearance: White to Pale Yellow/Powder
• Density: 2.17 g/cm³
• Vapor Pressure: 1.86E-23mmHg at 25°C
• Refractive Index: 1.876
• Storage Temp.: 2-8°C
• Solubility: DMF: 20 mg/mL, clear, faintly yellow
• PKA: 13.05±0.70(Predicted)
• Water Solubility: Soluble in DMF, DMSO, methanol or ethanol. Sparingly soluble in water
• Stability: Stable for 1 year from date of purchase as supplied. Solutions in DMSO may be stored at -20°C for up to 3 months.
• Merck: 13,4152
• BRN: 1225932
• CAS DataBase Reference: Fludarabine(CAS DataBase Reference)
• NIST Chemistry Reference: Fludarabine(21679-14-1)
• EPA Substance Registry System: Fludarabine(21679-14-1)

Safety Data

• Hazard Codes: Xn
• Statements: 23/24/25-36/37/38-39/23/24/25-39-22
• Safety Statements: 26-36/37-45-36
• WGK Germany: 3
• RTECS: AU6207000
• F: 10
• Hazardous Substances Data: 21679-14-1(Hazardous Substances Data)

Usage

Originator

Fludarabine,Union Pharmaceutical

Indications

Fludarabine (Fludara) is a fluorinated purine analogue of the antiviral agent vidarabine.The active metabolite, 2-fluoro-ara-adenosine triphosphate, inhibits various enzymes involved in DNA synthesis, including DNA polymerase-α, ribonucleotide reductase, and DNA primase. Unlike most antimetabolites, it is toxic to nonproliferating as well as dividing cells, primarily lymphocytes and lymphoid cancer cells. The drug is highly active in the treatment of chronic lymphocytic leukemia, with approximately 40% of patients achieving remissions after previous therapy with alkylating agents has failed. Activity is also seen in the low-grade lymphomas. The major side effect is myelosuppression, which contributes to fevers and infections in as many as half of treated patients. Nausea and vomiting are mild. Occasional neurotoxicity has been noted at higher doses, with agitation, confusion, and visual disturbances.

Preparation

Synthesis of fludarabine 1: 2-Fluoroadenine is reacted with 9-β-D-arabinosyl-uracile, taking water as a solvent. The reaction will take place in the presence of Enterobacter aerogenes. Fludarabine so formed is then treated with acetic anhydride to form the acetylderivative.The acetyl derivative is then crystallize to get back the pure Fludarabine.Synthesis of fludarabine 2: 2,4,5,6-tetraaminopyrimidine and formamide were cyclized together by heating to give 2,6-diaminopurine. acylation with acetic anhydride-pyridine complex gave 2,6-diacetamidopurine. then reacted with 2,3,5-tri-o-benzyl-d-arabinofuranosyl chloride to give compound (i). (i) was deacetylated to give compound (ii). (ii) in fluoroboronic acid-tetrahydrofuran, first nitrosated and then substituted to give compound (ili). in the presence of boron trichloride, the benzyl group was removed to give fludarabine. the yield was 17.5% in terms of 2,3,5-tri-o-benzyl-d-arabinofuranosyl chloride.

Therapeutic Function

Antineoplastic

Biological Activity

Purine analog that inhibits DNA synthesis. Exhibits antiproliferative activity (IC 50 = 1.54 μ M in RPMI cells) and triggers apoptosis through increasing Bax and decreasing Bid, XIAP and survivin expression. Displays anticancer activity against hematological malignancies in vivo .

Biochem/physiol Actions

Fludarabine (the 5′-phosphate) is a prodrug that is converted to F-ara-A, which enters cells and accumulates primarily as the 5′-triphosphate. F-ara-A interferes with DNA synthesis and repair and induces apoptosis of cancer cells. F-ara-A also strongly inhibits DNA methylation, particularly methylation of cytosine in CpG dinucleotide sequences.

Mode of action

Fludarabine is a fluorinated analogue of adenine that is relatively resistant to deamination by adenosine deaminase. Fludarabine phosphate is a prodrug that is rapidly dephosphorylated to 2-fluoro-ara-A and then phosphorylated intracellularly by deoxycytidine kinase to the active metabolite triphosphate 2-fluoro-ara-ATP. Fludarabine inhibits the DNA synthesis via inhibition of ribonucleotide reductase, DNA polymerase (α, δ, and ε), DNA primase, and DNA ligase. The action mechanism also is by partial inhibition of RNA polymerase II, causing reduction in protein synthesis. It is believed that effects on DNA, RNA, and protein synthesis contribute to the inhibition of cell growth, mostly by inhibition of DNA synthesis. Lymphocytes of CLL when exposed, in vitro, to the compound 2-fluoro-ara-A lead to extensive DNA fragmentation and apoptosis.

References

Ross et al. (1993), A review of its pharmacological properties and therapeutic potential in malignancy; Drugs, 45 737 Gandhi and Plunkett (2002), Cellular and clinical pharmacology of fludarabine; Pharmacokinet. 41 93 Langenhorst et al. (2019), Fludarabine exposure in the conditioning prior to allogeneic hematopoietic cell transplantation predicts outcomes; Blood Adv., 3 2179 Nishioka et al. (2008), Fludarabine induces growth arrest and apoptosis of cytokine- or alloantigen-stimulated peripheral blood mononuclear cells and decreases production of Th1 cytokines via inhibition of nuclear factor kappaB; Bone Marrow Transplant., 41 303 Jensen et al. (2012), Cytotoxic purine nucleoside analogues bind to A1, A2A and A3 adenosine receptors; Naunyn Schmiedebergs Arch. Pharmacol., 385 519

InChI:InChI=1/C10H12FN5O4/c11-10-14-7(12)4-8(15-10)16(2-13-4)9-6(19)5(18)3(1-17)20-9/h2-3,5-6,9,17-19H,1H2,(H2,12,14,15)/t3-,5-,6+,9-/m1/s1

Upstream product

• 3083-77-0 araU 
• 700-49-2 2-fluoroadenine 
• 24649-69-2 2-fluoro-9-(2,3,5-tri-o-benzyl-beta-D-arabinofuranosyl)adenine 
• 22224-41-5 1,3,5-tri-O-benzoyl-α-D-ribofuranoside 
• 38819-10-2 9-β-D-arabinofuranosylguanine 
• 5536-17-4 arabinosyl adenine 
• 118-00-3 G 
• 146-78-1 2-fluoroadenosine 
• 161109-77-9 2-fluoro-ara-adenine triacetate 
• 52522-49-3 2,3,5-tri-O-benzyl-1-(4-nitrobenzoyloxy)-D-arabinofuranose 
• 4060-34-8 2,3,5-tri-O-benzyl-α-D-arabinofuranosyl chloride 
• 143482-58-0 6-azido-2-fluoropurine 
• 75607-67-9 fludarabine phosphate 

Downstream Products

• 700-49-2 2-fluoroadenine 
• 72075-46-8 O¹-Phosphono-α-D-arabinofuranose 
• 75607-67-9 fludarabine phosphate 

Relevant articles and documents

Practical Synthesis of Fludarabine and Nelarabine

A new practical synthesis strategy has been developed to access the β- d -arabinofuranosyl purine nucleosides fludarabine and nelarabine. In our approach, an ortho -alkyne benzoyl ester is transiently introduced as a neighbouring-participation group in Vorbrüggen glycosylation to afford the corresponding β-nucleoside exclusively. The latter was further removed efficiently by using freshly prepared Ph 3 PAuOTFA to give the corresponding 2′-OH nucleosides without transesterification. After reversion of the configuration of 2′-OH and deprotection, fludarabine and nelarabine were obtained in high yield and purity.

Synthesis method of fludarabine phosphate

The invention provides a synthesis method of fludarabine phosphate, and the synthesis route is as follows: with vidarabine as a starting raw material, fludarabine is obtained through upper protection,nitrification, fluorination denitration and deprotection, so that the fludarabine is prepared by adopting a brand-new synthesis route; meanwhile, by improving the process of phosphorylation and refining of fludarabine, the reaction time is shortened, the generation of by-products is reduced, and the product quality is improved. The method has the following advantages: 1, the initial raw materialis beta-configuration, isomer separation is avoided, and the yield is improved; raw materials are easy to obtain, the route is simple, and the price is low; 3, salification and column-passing purification and separation are avoided, so that the method is suitable for industrialization.

Synthesis method of fludarabine and nelarabine

The present invention discloses a synthesis method of fludarabine and nelarabine. The method is prepared from a ribofuranose derivative as a raw material, an o-alkynyl benzoate is introduced at a 2-position hydroxyl group to obtain a key glycosyl donor, the key glycosyl donor and purine bases are subjected to a coupling reaction using a Vorbruggen glycosylation reaction to highly efficiently construct beta-nucleoside bonds, under catalysis of a gold catalyst, 2'-position ester groups are selectively removed to obtain important furyl ribonucleotide, and a bare 2' hydroxyl group is then subjected to two-step reactions of hydroxyl inversion and deprotection to respectively obtain fludarabine and nelarabine. The synthesis strategy has characteristics of being simple in operation, simple and easy to obtain the raw materials, easy in separation of products, high in reaction yield, etc., and has relatively good prospects for popularization and application.

Enzymatic Synthesis of Therapeutic Nucleosides using a Highly Versatile Purine Nucleoside 2’-DeoxyribosylTransferase from Trypanosoma brucei

The use of enzymes for the synthesis of nucleoside analogues offers several advantages over multistep chemical methods, including chemo-, regio- and stereoselectivity as well as milder reaction conditions. Herein, the production, characterization and utilization of a purine nucleoside 2’-deoxyribosyltransferase (PDT) from Trypanosoma brucei are reported. TbPDT is a dimer which displays not only excellent activity and stability over a broad range of temperatures (50–70 °C), pH (4–7) and ionic strength (0–500 mM NaCl) but also an unusual high stability under alkaline conditions (pH 8–10). TbPDT is shown to be proficient in the biosynthesis of numerous therapeutic nucleosides, including didanosine, vidarabine, cladribine, fludarabine and nelarabine. The structure-guided replacement of Val11 with either Ala or Ser resulted in variants with 2.8-fold greater activity. TbPDT was also covalently immobilized on glutaraldehyde-activated magnetic microspheres. MTbPDT3 was selected as the best derivative (4200 IU/g, activity recovery of 22 %), and could be easily recaptured and recycled for >25 reactions with negligible loss of activity. Finally, MTbPDT3 was successfully employed in the expedient synthesis of several nucleoside analogues. Taken together, our results support the notion that TbPDT has good potential as an industrial biocatalyst for the synthesis of a wide range of therapeutic nucleosides through an efficient and environmentally friendly methodology.

Synthesis process of fludarabine base

The invention provides a synthesis process of fludarabine base. The synthesis process comprises selecting raw materials, synthesizing an intermediate, and synthesizing fludarabine base. The synthesisprocess has the beneficial effects that the source of the raw materials is widely available, the cost of the raw materials is low, the synthesis process is suitable for large-scale production and hassimple synthesis steps, fewer harmful substances are discharged in synthesis, and the synthesis process is safe and environment-friendly, has good use effect and can be promoted.

The arsenolysis reaction in the biotechnological method of synthesis of modified purine β-D-arabinonucleosides

We found a unique property of E. coli purine nucleoside phosphorylases to selectively perform the arsenolysis reaction of ribonucleosides in their active site without affecting β-D-arabinonucleosides. In the synthesis of modified β-D-arabinonucleosides from the corresponding ribonucleosides, the catalytical amount of sodium arsenate in the transglycosylation reaction provided a 95 to 98% conversion rate. Such an approach was shown to simplify the composition of the reaction mixtures and facilitate the isolation of the target nucleosides, particularly, vidarabine, fludarabine, and nelarabine.

Simple modification to obtain high quality fludarabine

A simple and improved debenzylation process is described to obtain fludarabine in greater than 99.8% purity and 90-95% yield.

Use of Citrobacter koseri whole cells for the production of arabinonucleosides: A larger scale approach

Purine arabinosides are well known antiviral and antineoplastic drugs. Since their chemical synthesis is complex, time-consuming, and polluting, enzymatic synthesis provides an advantageous alternative. In this work, we describe the microbial whole cell synthesis of purine arabinosides through nucleoside phosphorylase-catalyzed transglycosylation starting from their pyrimidine precursors. By screening of our microbial collection, Citrobacter koseri (CECT 856) was selected as the best biocatalyst for the proposed biotransformation. In order to enlarge the scale of the transformations to 150 mL for future industrial applications, the biocatalyst immobilization by entrapment techniques and its behavior in different reactor configurations, considering both batch and continuous processes, were analyzed. C. koseri immobilized in agarose could be used up to 68 times and the storage stability was at least 9 months. By this approach, fludarabine (58% yield in 14 h), vidarabine (71% yield in 26 h) and 2,6-diaminopurine arabinoside (77% yield in 24 h), were prepared.

THERAPEUTIC FOR HEPATIC CANCER

A novel pharmaceutical composition for treating or preventing hepatocellular carcinoma and a method of treatment are provided. A pharmaceutical composition for treating or preventing liver cancer is obtained by combining a chemotherapeutic agent with an anti-glypican 3 antibody. Also disclosed is a pharmaceutical composition for treating or preventing liver cancer which comprises as an active ingredient an anti-glypican 3 antibody for use in combination with a chemotherapeutic agent, or which comprises as an active ingredient a chemotherapeutic agent for use in combination with an anti-glypican 3 antibody. Using the chemotherapeutic agent and the anti-glypican 3 antibody in combination yields better therapeutic effects than using the chemotherapeutic agent alone, and mitigates side effects that arise from liver cancer treatment with the chemotherapeutic agent.

METHOD FOR THE MANUFACTURE OF 2-FLUORO-ARA-ADENINE

A method is described for the manufacture of pure 2-fluoro-ara-adenine of Formula (I) from 2-fluoro-ara-adenine triacetate using potassium carbonate (K2CO3), wherein the 2-fluoro-ara-adenine has a reduced dimer contents, as well as the compound 2-fluoro-ara-adenine having a dimer contents of ≤ 0,3 %.