Daratumumab (Synonyms: Daremuzumab; Anti-Human CD38, Human Antibody)

Cell lines

bone marrow mononuclear cells (BM-MNC)

Preparation Method

Freshly isolated BM-MNCs, containing 2% to 35% malignant plasma cells as determined by flow cytometry, were immediately used in ex vivo experiments. The BM-MNCs, containing the malignant plasma cells, as well as the patient's own effector cells, were incubated with daratumumab (10 µg/mL), lenalidomide (3 µmol/L), and bortezomib (3 nmol/L) alone or in combination in RPMI + 10% FBS in 96-well round bottom plates in fully humified incubators at 37°C, 5% CO2-air mixture for 48 hours. The survival of primary CD138+ multiple myeloma cells was determined by flow cytometry.

Reaction Conditions

10 µg/mL for 48 hours

Applications

PF-07104091 inhibited T47D and KPL1 cells with IC50s of 0.785 and 0.603 µM, respectively.

Animal models

human multiple myeloma model in the RAG2-/-γc-/- mice

Preparation Method

A humanized microenvironment was generated in mice by subcutaneous implantation of ceramic scaffolds that were seeded with human MSC (2 × 105 cells/scaffold) and in vitro cultured for 7 days in osteogenic medium, containing ascorbic acid and dexamethasone. Eight weeks after implantation, mice received a sublethal irradiation dose (3 Gy, 200 kV, 4 mA) and luciferase-gene-marked primary multiple myeloma cells were injected directly into the scaffolds (1 × 106 cells/scaffold). Luciferase transduction of primary multiple myeloma cells was carried out using the lentiviral construct pRRL-cPPT-CMV-Luc2-IRES-GFP-PRE-SIN. When tumors became clearly detectable, mice were distributed over the following treatment groups: (i) control, (ii) T-cell depleted PBMC (PBMC-T), (iii) PBMC-T plus lenalidomide, (iv) PBMC-T plus daratumumab, and (v) PBMC-T plus lenalidomide plus daratumumab. Lenalidomide (1 mg/kg) was given in 5 days on 2 days off schedule for 2 weeks (days 49-53 and 56-60) and both daratumumab (8 mg/kg) and PBMC-T (8 × 106 cells/mouse) were given on days 49 and 56.

Dosage form

8 mg/kg on days 49 and 56.

Applications

Treatment with daratumumab alone suppressed the tumor growth significantly. And the combination of daratumumab plus lenalidomide was able to reduce the tumor volume.

References:

[1]: Nijhof IS, Groen RW, Noort WA, van Kessel B, de Jong-Korlaar R, Bakker J, Van Bueren JJ, Parren PW, Lokhorst HM, Van De Donk NW, Martens AC. Preclinical evidence for the therapeutic potential of CD38-targeted immuno-chemotherapy in multiple myeloma patients refractory to lenalidomide and bortezomib. Clinical cancer research. 2015 Jun 15;21(12):2802-10.