Cladribine (also known as 2-CdA, 2-chlorodeoxyadenosine; Leustatin; Leustat; Leustatine), a purine nucleoside antimetabolite analogue and an approved medication used for treating hairy cell leukemia and B-cell chronic lymphocytic leukemia, is an adenosine deaminase inhibitor for U266, RPMI8226, and MM1.S cells with IC50s of approximately 2.43 μM, 0.75 μM, and 0.18 μM, respectively. It is a well-known purine nucleoside analog that exhibits unique anti-lymphoma-proliferative activity against diseases like hairy cell leukemia (HCL). Cladribine exhibited dose-dependent inhibition of cell proliferation in U266, RPMI8226, and MM1.S cells, with IC50 values of 2.43, 0.75, and 0.18 μmol/L, in that order.
Physicochemical Properties
| Molecular Formula | C10H12CLN5O3 | |
| Molecular Weight | 285.69 | |
| Exact Mass | 285.062 | |
| Elemental Analysis | C, 42.04; H, 4.23; Cl, 12.41; N, 24.51; O, 16.80 | |
| CAS # | 4291-63-8 | |
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| PubChem CID | 20279 | |
| Appearance | White to off-white solid powder | |
| Density | 2.0±0.1 g/cm3 | |
| Boiling Point | 547.6±60.0 °C at 760 mmHg | |
| Melting Point | 181-185 °C(lit.) | |
| Flash Point | 285.0±32.9 °C | |
| Vapour Pressure | 0.0±1.5 mmHg at 25°C | |
| Index of Refraction | 1.871 | |
| LogP | 0.02 | |
| Hydrogen Bond Donor Count | 3 | |
| Hydrogen Bond Acceptor Count | 7 | |
| Rotatable Bond Count | 2 | |
| Heavy Atom Count | 19 | |
| Complexity | 338 | |
| Defined Atom Stereocenter Count | 3 | |
| SMILES | ClC1=NC(=C2C(=N1)N(C([H])=N2)[C@@]1([H])C([H])([H])[C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])N([H])[H] |
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| InChi Key | PTOAARAWEBMLNO-KVQBGUIXSA-N | |
| InChi Code | InChI=1S/C10H12ClN5O3/c11-10-14-8(12)7-9(15-10)16(3-13-7)6-1-4(18)5(2-17)19-6/h3-6,17-18H,1-2H2,(H2,12,14,15)/t4-,5+,6+/m0/s1 | |
| Chemical Name | (2R,3S,5R)-5-(6-amino-2-chloropurin-9-yl)-2-(hydroxymethyl)oxolan-3-ol | |
| Synonyms | 2-Chloro-2′-deoxyadenosine; CldAdo; 2CdA; 2-CdA, 2-chlorodeoxyadenosine; Cladribina. Trade name: Leustatin; Leustat; 2-Chloro-2'-deoxyadenosine; 4291-63-8; Leustatin; 2-Chlorodeoxyadenosine; 2-CdA; Chlorodeoxyadenosine; Litak; Leustatine | |
| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
Adenosine deaminase (MM1.S cells) ( IC50 = 0.18 μM ) Adenosine deaminase (RPMI8226 cells) ( IC50 = 0.75 μM ) Adenosine deaminase (U266 cells) ( IC50 = 2.43 μM ) |
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| Cell Assay | The viability of the cells is assessed using the non-radioactive cell proliferation kit. To summarize, 0.1 mL of complete medium (5% FBS) is used as a control, or 0.1 mL of the same medium containing multiple doses of cladribine, is added to 96-well plates, and the cells are incubated for 72 hours. Reduction of MTS is used to calculate the percentages of surviving cells from each group in comparison to controls, which is defined as 100% survival, after reading all wells at 490 nm using a micro-plate reader. | ||
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| ADME/Pharmacokinetics |
Absorption, Distribution and Excretion Oral bioavailability is 34 to 48%. 4.5 ± 2.8 L/kg [patients with hematologic malignancies] 9 L/kg 978 +/- 422 mL/h/kg Cladribine is bound approximately 20% to plasma proteins. It is not known whether cladribine is distributed into breast milk. It is not known whether cladribine is removed from circulation by dialysis or hemofiltration. Cladribine penetrates into cerebrospinal fluid. One report indicates that concentrations are approximately 25% of those in plasma. For more Absorption, Distribution and Excretion (Complete) data for CLADRIBINE (9 total), please visit the HSDB record page. Metabolism / Metabolites Metabolized in all cells with deoxycytidine kinase activity to 2-chloro-2'-deoxyadenosine-5'-triphosphate Metabolized in all cells with deoxycytidine kinase activity to 2-chloro-2'-deoxyadenosine-5'-triphosphate. Metabolized in all cells with deoxycytidine kinase activity to 2-chloro-2'-deoxyadenosine-5'-triphosphate Half Life: 5.4 hours Biological Half-Life 5.4 hours Cladribine plasma concentration after intravenous administration declines multi-exponentially with an average half-life of 6.7 +/-2.5 hours. ... The terminal half-life varies from 5.7 to 19.7 hours ... ... The terminal phase half-life in 22 patients ranged from 14.3-25.8 hr, with a mean (SD) of 19.7 (3.4) hr. ... |
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| Toxicity/Toxicokinetics |
Hepatotoxicity In clinical trials, cladribine was not associated with elevations in serum enzymes or bilirubin levels either during or after therapy. Since its approval and wide scale use in hairy cell leukemia there have been no reports of clinically apparent liver injury attributable to cladribine administration. Likelihood score: E (unlikely cause of clinically apparent liver injury). Effects During Pregnancy and Lactation ◉ Summary of Use during Lactation The amount of cladribine in milk is low with oral doses of 10 to 20 mg daily used in multiple sclerosis. Data in one patient indicates that the drug is rapidly eliminated over 24 hours and undetectable at 48 hours after a dose. Manufacturers recommend a 7-day (Europe) or 10-day (US) abstinence period. Chemotherapy may adversely affect the normal microbiome and chemical makeup of breastmilk. Women who receive chemotherapy during pregnancy are more likely to have difficulty nursing their infant. ◉ Effects in Breastfed Infants Relevant published information was not found as of the revision date. ◉ Effects on Lactation and Breastmilk Relevant published information was not found as of the revision date. Protein Binding 20% |
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| References |
[1]. BMC Cancer . 2011 Jun 16:11:255. [2]. Cancer Chemother Pharmacol . 1998;42(1):77-83. [3]. Exp Hematol . 2010 Sep;38(9):744-55. [4]. Eur J Neurol . 2009 Mar;16(3):409-12. [5]. Biomarkers . 2009 Dec;14(8):554-9. [6]. Drug Metabol Drug Interact . 2008;23(3-4):291-8. [7]. Ann Hematol . 2002 Sep;81(9):508-13. |
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| Additional Infomation |
Cladribine can cause developmental toxicity according to state or federal government labeling requirements. Cladribine is 2'-Deoxyadenosine in which the hydrogen at position 2 on the purine ring has been substituted by chlorine. It inhibits the synthesis and repair of DNA, particularly in lymphocytes and monocytes, and is used as an antimetabolite antineoplastic drug for the treatment of lymphoid malignancies including hairy-cell leukaemia and chronic lymphocytic leukaemia. It has a role as an antineoplastic agent and an immunosuppressive agent. It is a purine 2'-deoxyribonucleoside and an organochlorine compound. An antineoplastic agent used in the treatment of lymphoproliferative diseases including hairy-cell leukemia. Cladribine is a Purine Antimetabolite. Cladribine is a purine analogue and antineoplastic agent used primarily in the therapy of hairy cell leukemia. Cladribine is typically given intravenously daily for 7 days, usually as a single course, and has not been associated with serum enzyme elevations during therapy or with instances of clinically apparent acute liver injury with jaundice. Cladribine is a purine nucleoside antimetabolite analogue. Cladribine triphosphate, a phosphorylated metabolite of cladribine, incorporates into DNA, resulting in single-strand breaks in DNA, depletion of nicotinamide adenine dinucleotide (NAD) and adenosine triphosphate (ATP), and apoptosis. Because this agent is resistant to adenosine deaminase, an enzyme that inactivates some antineoplastic agents, it is selectively toxic to lymphocytes and monocytes which exhibit little deoxynucleotide deaminase activity. (NCI04) An antineoplastic agent used in the treatment of lymphoproliferative diseases including hairy-cell leukemia. An antineoplastic agent used in the treatment of lymphoproliferative diseases including hairy-cell leukemia. Drug Indication For the treatment of active hairy cell leukemia (leukemic reticuloendotheliosis) as defined by clinically significant anemia, neutropenia, thrombocytopenia, or disease-related symptoms. Also used as an alternative agent for the treatment of chronic lymphocytic leukemia (CLL), low-grade non-Hodgkin's lymphoma, and cutaneous T-cell lymphoma. Treatment of adult patients with highly active relapsing multiple sclerosis (MS) as defined by clinical or imaging features. Litak is indicated for the treatment of hairy-cell leukaemia. Multiple Sclerosis Mechanism of Action Cladribine is structurally related to fludarabine and pentostatin but has a different mechanism of action. Although the exact mechanism of action has not been fully determined, evidence shows that cladribine is phosphorylated by deoxycytidine kinase to the nucleotidecladribine triphosphate (CdATP; 2-chloro-2′-deoxyadenosine 5′-triphosphate), which accumulates and is incorporated into DNA in cells such as lymphocytes that contain high levels of deoxycytidine kinase and low levels of deoxynucleotidase, resulting in DNA strand breakage and inhibition of DNA synthesis and repair. High levels of CdATP also appear to inhibit ribonucleotide reductase, which leads to an imbalance in triphosphorylated deoxynucleotide (dNTP) pools and subsequent DNA strand breaks, inhibition of DNA synthesis and repair, nicotinamide adenine dinucleotide (NAD) and ATP depletion, and cell death. Unlike other antimetabolite drugs, cladribine has cytotoxic effects on resting as well as proliferating lymphocytes. However, it does cause cells to accumulate at the G1/S phase junction, suggesting that cytotoxicity is associated with events critical to cell entry into S phase. It also binds purine nucleoside phosphorylase (PNP), however no relationship between this binding and a mechanism of action has been established. Cladribine is an antimetabolite. The exact mechanism of action in hairy cell leukemia is unknown. Cladribine is resistant to the action of adenosine deaminase (ADA), which deaminates deoxyadenosine to deoxyinosine. The phosphorylated metabolites of cladribine accumulate in cells with a high ratio of deoxycytidine kinase activity to 5' nucleotidase activity (lymphocytes, monocytes ) and are converted to the active triphosphate deoxynucleotide. Intracellular accumulation of toxic deoxynucleotides selectively kills these cells, which become unable to properly repair single-strand DNA breaks, leading to disruption of cell metabolism. In addition, there is some evidence that deoxynucleotides are incorporated into the DNA of dividing cells and impair DNA synthesis. Cladribine also induces apoptosis (a form of programmed cell death in sensitive cells). Cladribine's action is cell cycle-phase nonspecific; cladribine equally affects dividing and resting lymphocytes. Cladribine has immunosuppressant activity ; restoration of lymphocyte subsets after treatment takes at least 6 to 12 months, although clinical immunocompetence is usually restored after about a month. Significant reductions in T and B lymphocytes occur during treatment (both CD4 and CD8 are affected) and CD4 counts recover more slowly after treatment. /Investigators/ have studied the role of caspases and mitochondria in apoptosis induced by 2-chloro-2'-deoxyadenosine (cladribine) in several human leukemic cell lines. Cladribine treatment induced mitochondrial transmembrane potential (DeltaPsi(m)) loss, phosphatidylserine exposure, caspase activation and development of typical apoptotic morphology in JM1 (pre-B), Jurkat (T) and U937 (promonocytic) cells. Western-blot analysis of cell extracts revealed the activation of at least caspases 3, 6, 8 and 9. Co-treatment with Z-VAD-fmk (benzyloxy-carbonyl-Val-Ala-Asp-fluoromethylketone), a general caspase inhibitor, significantly prevented cladribine-induced death in JM1 and Jurkat cells for the first approximately 40 h, but not for longer times. Z-VAD-fmk also partly prevented some morphological and biochemical features of apoptosis in U937 cells, but not cell death. Co-incubation with selective caspase inhibitors Ac-DEVD-CHO (N-acetyl-Asp-Glu-Val-Asp-aldehyde), Ac-LEHD-CHO (N-acetyl-Leu-Glu-His-Asp-aldehyde) or Z-IETD-fmk (benzyloxycarbonyl-Ile-Glu-Thr-Asp-fluoromethylketone), inhibition of protein synthesis with cycloheximide or cell-cycle arrest with aphidicolin did not prevent cell death. Overexpression of Bcl-2, but not CrmA, efficiently prevented death in Jurkat cells. In all cell lines, death was always preceded by Delta Psi(m) loss and accompanied by the translocation of the protein apoptosis-inducing factor (AIF) from mitochondria to the nucleus. These results suggest that caspases are differentially involved in induction and execution of apoptosis depending on the leukemic cell lineage. In any case, Delta Psi(m) loss marked the point of no return in apoptosis and may be caused by two different pathways, one caspase-dependent and the other caspase-independent. Execution of apoptosis was always performed after Delta Psi(m) loss by a caspase-9-triggered caspase cascade and the action of AIF. Cladribine (chlorodeoxyadenosine, 2-CdA), a synthetic purine nucleoside, is an antineoplastic agent. ... The precise mechanism(s) of antileukemic action of cladribine has not been fully elucidated. Cladribine is phosphorylated by deoxycytidine kinase to the nucleotide cladribine triphosphate (CdATP; 2-chloro-2'-deoxyadenosine 5'-triphosphate), which accumulates and is incorporated into DNA in cells such as lymphocytes that have high levels of deoxycytidine kinase and low levels of deoxynucleotidase. High intracellular concentrations of cladribine triphosphate appear to inhibit ribonucleotide reductase, causing an imbalance in triphosphorylated deoxynucleotide (dNTP) pools and subsequent DNA strand breaks, inhibition of DNA synthesis and repair, nicotinamide adenine dinucleotide (NAD) and ATP depletion, and cell death. Incorporation of accumulated cladribine triphosphate into DNA also may contribute to DNA strand breakage and inhibition of DNA synthesis and repair. Unlike other commonly used antineoplastic drugs that affect purine and pyrimidine metabolism, cladribine has cytotoxic effects on resting as well as proliferating lymphocytes and monocytes. |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: 25 mg/mL (87.51 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication. Solubility in Formulation 2: 5% DMSO+ 30% PEG 300+ 1% Tween 80+ H2O: 10mg/mL (35.00mM) (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.5003 mL | 17.5015 mL | 35.0030 mL | |
| 5 mM | 0.7001 mL | 3.5003 mL | 7.0006 mL | |
| 10 mM | 0.3500 mL | 1.7501 mL | 3.5003 mL |