Lenalidomide

Biological Activity

Lenalidomide (Revlimid) is a derivative of thalidomide. Lenalidomide has tumoricidal and immunomodulatory activity against multiple myeloma. Lenalidomide has also shown efficacy in the class of hematological disorders known as myelodysplastic syndromes (MDS). Lenalidomide maintenance therapy, initiated at day 100 after hematopoietic stem-cell transplantation, was associated with more toxicity and second cancers. Lenalidomide has significantly improved overall survival in myeloma (which generally carries a poor prognosis), although toxicity remains an issue for users.

Product Citations

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  Blood. 2021 Dec 23;138(25):2655-2669.

  Targeting intracellular WT1 in AML with a novel RMF-peptide-MHC specific T-cell bispecific antibody
  Lenalidomide purchased from AbMole

• 

  BMC Biol. 2021 May 20;19(1):108.

  Very long intergenic non-coding (vlinc) RNAs directly regulate multiple genes in cis and trans
  Lenalidomide purchased from AbMole

• 

  Int Immunopharmacol. 2015 Sep;578-87.

  Resveratrol attenuates hypoxia-induced neurotoxicity through inhibiting microglial activation.
  Lenalidomide purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Int Immunopharmacol (2015). Lenalidomide, Figure 9. (AbMole BioScience, Houston, TX, USA)
	Method	MTT assay
	Cell Lines	BV-2 cells
	Concentrations	10 μM
	Incubation Time	30 min
	Results	Lenalidomide significantly decreased hypoxia-induced mRNA levels of TNF-alpha (Fig. 9A). Next to determine whether the inhibition of TNF-alpha contributes to microglial-CM induced neuron death, BV-2 cells were pretreated with lenalidomide (10 μM) for 30 min, and then exposed hypoxia for 24 h; neuronal cells were treated with different CM from BV-2 cells for 24 h. MTT assay showed that the viability of neurons in the hypoxia-CM condition was significantly decreased, while this effect was reversed by the CM treated with hypoxia and lenalidomide (Fig. 9B).

	Source	Graduation Thesis (2015) . Figure 14.Lonidamine (AbMole BioScience, CAS No.: 50264-69-2)
	Method	hochest staining and Annexin-V/PI flow cytometry
	Cell Lines	HL60, MOLM13 and NB4 cells
	Concentrations	1, 5, 20, 50, 100 µM
	Incubation Time	48 h
	Results	An overall higher degree of reduced metabolic activity was observed from three independent studies with MOLM13 (Figure 14A) and NB4 (Figure 14B) cells thus were suggested being more responsive to co-treatment.

	Source	Graduation Thesis (2015) . Figure 13.Lonidamine (AbMole BioScience, CAS No.: 50264-69-2)
	Method	hochest staining and Annexin-V/PI flow cytometry
	Cell Lines	HL60, MOLM13 and NB4 cells
	Concentrations	1, 5, 20, 50, 100 µM
	Incubation Time	48 h
	Results	These data indicated a comparable trend of viability in all three cell lines following combinatorial treatment of VPA and LND (Figure 13B, C). HL60 cells are only presented for Hoechst viability scoring in figure 13A since the cells do not expose phosphatidylserine upon apoptosis induction.

	Source	Graduation Thesis (2015) . Figure 12.Lonidamine (AbMole BioScience, CAS No.: 50264-69-2)
	Method	WST-1 proliferation based assay
	Cell Lines	HL60 cells
	Concentrations	50, 100 µM
	Incubation Time	48 h
	Results	Maximal potentiation of LND was obtained by 100 µM LND in the sequence VPA→LND+VPA, but no significant increased anti-proliferative effect was obtained from three independent and replicable studies.

	Source	Graduation Thesis (2015) . Figure 11.Lonidamine (AbMole BioScience, CAS No.: 50264-69-2)
	Method	Hochest staining
	Cell Lines	HL60, MOLM13 and NB4 cells
	Concentrations	1 µM ~ 500 µM
	Incubation Time	24 h and 48 h
	Results	The frequencies of abnormal nucleic morphology were minor at concentrations lower than 50 µM in all three cell lines.

	Source	Graduation Thesis (2015) . Figure 10.Lonidamine (AbMole BioScience, CAS No.: 50264-69-2)
	Method	WST-1 proliferation based assay
	Cell Lines	HL60, MOLM13 and NB4 cells
	Concentrations	20 µM ~ 500 µM
	Incubation Time	48 h
	Results	The IC50 of LND in NB4 (201 µM), HL60 (248 µM) and MOLM13 (124 µM) are presented in Figure 10 B, C and D indicating LND to be more effective after 48 hour treatment in MOLM13 and NB4 cells at lower concentrations than when treated for 24 hours.

	Source	Graduation Thesis (2015) . Figure 9.Lonidamine (AbMole BioScience, CAS No.: 50264-69-2)
	Method	WST-1 proliferation based assay
	Cell Lines	HL60, MOLM13 and NB4 cells
	Concentrations	HL60 (149 µM, CI: 110-200 µM), MOLM13 (218 µM, CI: 153 – 311 µM), and NB4 (203µM, CI: 106 – 387 µM)
	Incubation Time	24 h
	Results	IC50s of LND was obtained from WST-1 proliferation assay, and are presented in Figure 9B, C and D for HL60 (149 µM), MOLM13 (218 µM) and NB4 (203 µM), respectively.

	Source	Graduation Thesis (2015) . Figure 7.Lonidamine (AbMole BioScience, CAS No.: 50264-69-2)
	Method	WST-1 proliferation based assay
	Cell Lines	NB4 cells
	Concentrations	12.5, 25, 50, 75, 100 µ M
	Incubation Time	48 h
	Results	the combination of LND and MMF neither showed significant enhanced potentiation of MMF nor LND.

Protocol (for reference only)

Cell Experiment
Cell lines	HL60, MOLM13 and NB4 cells
Preparation method	Spectrophotometric quantification of cell proliferation was assayed by the water soluble tetrazolium salt-1 (WST-1) (Roche Ltd, Basel, Switzerland) based colorimetric assay. This tetrazolium salt is reduced extracellularly to formazan dye by enzymes of the plasma membrane oxidoreductase. (122) The primary reductant is NADH derived from the TCA of the mitochondria. Hence, WST-1 is converted by metabolically active cells and was employed to measure cell proliferation. The cell lines HL60, MOLM13 and NB4 were plated in triplicate (20×103 cells/well) in a 96-well microplate and treated with either LND (1, 5, 20, 50, 100, 200, and 500 µM) .
Concentrations	1, 5, 20, 50, 100, 200, and 500 µM
Incubation time	48 h

Animal Experiment
Animal models	NOD/scid/gamma (NSG) mice
Formulation	10% DMSO, 70% PEG300
Dosages	50 mg/kg daily
Administration	i.p.

Chemical Information

Molecular Weight	259.26
Formula	C13H13N3O3
CAS Number	191732-72-6
Solubility (25°C)	DMSO 34 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] McCarthy PL, et al. N Engl J Med. Lenalidomide after stem-cell transplantation for multiple myeloma.

[2] Palumbo A, et al. N Engl J Med. Continuous lenalidomide treatment for newly diagnosed multiple myeloma.