Carbamoylphosphonates inhibit autotaxin and metastasis formation in vivo

Figures & data

Table 1. Inhibition constants of CPOs on MMP 2, CA IX and XII and ATX enzymes.*†.

		CPO, n=
		5	6	7	8
MMP	2	4	4	>100	>100
CA	IX	6.3	7	0.86	0.68
	XII	5.7	6.2	1.4	0.78
ATX		>100	1.5	1.5	1.5

*IC₅₀ units are µM.

†MMP-9 IC₅₀ > 100 for CPO n = 5–8.

Figure 1. The fluorescent image of the experimental animals at the different time points. ATX activity in vivo was estimated by the using ATX-Red-2, an in-vivo imaging probe, at a final dose of 0.5 mg/kg, for the visualization of enzymatic activity. The data were collected at three time points as indicated. Circle shows the tumor location.

Figure 2. The fluorescence ratio of the tumor versus the whole body fluorescence, as a function of time. There is a gradual decrease in the amount of fluorescence due to dye clearance from the mice, as time progresses. Dark grey bars represent the control mice while the light grey bars represent the treated ones.

Figure 3. The ATX activity as function of time in the treated mice. It is important to point out that ATX activity is still inhibited over 60% at 48 h following the administration of a single dose of the drug.

Table 2. In vivo effect of selected CPO inhibitors on metastasis formation in mice.* 

			Dose
			12.5 mg/kg	25 mg/kg
CPO Inhibitor symbol	Mode of introduction	CH2 chain length n	% decrease in metastases relative to control
JS-403	IP	6	82	82
JS-403	PO	6	61	71
TCH-18	PO	7	67	90

*Mice were injected IV with B16F10 cells and treated daily for three weeks by the drugs; values reflect percent decrease in lung metastases relative to control.

Table 3. Carbamoylphosphonates absorption to (A), and resorption from (B) hydroxyapatite (HAP).

Time (h)	CPO n = 5	CPO n = 6	CPO n = 7
Absorption (A) of CPOs to HAP (%)
0	0	0	0
1	33	37	57
2	35	49	59
18	41	51	67
Resorption (B) of CPOs from HAP (%)*
0	0	0	0
1	33	29	14
2	36	33	16
18	39	37	18
∞	100	100	100

*Samples for the resorption studies were taken from the 2 h absorption experiments.