Therapeutic efficacy of an injectable formulation of purinostat mesylate in SU-DHL-6 tumour model

Figures & data

Table 1. The gradient detection method of HPLC.

Time (minutes)	Mobile phase A (%)	Mobile phase B (%)
0	72	28
15	72	28
25	45	55
30	45	55
31	72	28
40	72	28

Table 2. The doses and schedules of the agents of the Hyper-CVAD + RTX group.

Drug	Dose(mg/kg)	Route	Schedule
Cyclophosphamide	10	i.p.	Administrated once on the first day of the 21-day cycles
Vincristine Sulphate	0.125	i.v.	Administrated once on the first day of the 21-day cycles
Doxorubicin Hydrochloride	0.825	i.v.	Administrated once on the first day of the 21-day cycles
Dexamethasone	1	i.p.	Administrated at day 1, day3, day5, day8 and day 12 of the 21-day cycles
Methotrexate	10	i.p.	Administrated at day2, day5, day9 and day12 of the 21-day cycles
Cytarabine	75	i.p.	Administrated at day3, day6, day10 and day13 of the 21-day cycles
Rituximab（RTX）	10	i.p.	BIW (twice per week)

Figure 1. Phase solubility diagram of PM/HP-β-CD system.

Figure 2. Computer molecular simulation of PM and HP-β-CD.

Figure 3. FT-IR spectra of PM, HP-β-CD, PM/HP-β-CD and the physical mixture (1:1 molar ratio).

Figure 4. Scanning electron microphotographs of PM, HP-β-CD, the physical mixture (1:1 molar ratio), and PM/HP-β-CD.

Figure 5. 1H-NMR spectra and chemical structural formula of PM, HP-β-CD, PM/HP-β-CD and the physical mixture (1:1 molar ratio).

Table 3. Chemical shift (δ, ppm) change values relating to the signals of PM in different states.

Site	PM	Physical mixture	PM/HP-β-CD	Δ δ
18, 20	8.73	8.72	8.71	−0.02
26, 30	8.36	8.34	8.10	−0.26
27, 29	7.17	7.12	6.60	−0.57

Figure 6. In vitro antitumour effect. (A) Free PM and PM/HP-β-CD induced apoptosis in SU-DHL-6 cells. (B) A histogram of apoptosis data. (C) Cell viability of free PM and PM/HP-β-CD on SU-DHL-6 cells.

Figure 7. In vivo anti-tumour effect in SU-DHL-6 xenograft subcutaneous tumour model. (A) Growth curve of the tumour. (B) Bodyweight of the mice in each group. (C) Tumour weight of each group. (D) Survival curve of each group during survival investigation. (E) Tumour growth curves in each group during survival investigation. (F) Photographs of subcutaneous tumours in each group.

Table 4. Antitumour effects of PM/HP-β-CD in human double-hit lymphoma SU-DHL-6 xenograft tumours.

Group	Dose mg/kg	Number	Body weight (g) X ± SD		Tumour volume (mm^3)			RTV	Tumour weight (g)
		End/ begin			X ± SD		T/C%
			Begin	End	Begin	End			X ± SD	Inhibition (%)
Control	–	7/7	22.30 ± 0.4	27.3 ± 1.7	144.9 ± 19.1	3651.3 ± 811.7	–	26.06	4.92 ± 1.05	–
Vehicle	–	7/7	22.60 ± 1.0	26.4 ± 2.2	148.1 ± 19.1	4146.1 ± 1047.3	108.63	28.31	4.99 ± 1.49	−1.33
LBH589	–	7/7	22.4 ± 0.5	26.4 ± 1.8	152.7 ± 30.6	4015.5 ± 1048.0	101.83	26.54	3.81 ± 1.08	22.72
HyperCVAD	–	7/7	22.4 ± 0.5	26.2 ± 1.1	154.7 ± 36.9	3276.8 ± 981.2	82.03	21.38	3.44 ± 1.23^a	30.08
RTX	10	7/7	22.4 ± 0.4	25.1 ± 1.0	153.7 ± 33.1	3505.3 ± 917.1	92.71	24.16	3.47 ± 1.07^ad	29.47
HyperCVAD + RTX	–	7/7	22.5 ± 0.6	25.1 ± 1.8	155.2 ± 38.1	3425.4 ± 1347.2	83.29	21.71	3.29 ± 1.12^ad	33.28
PM	2.5	7/7	22.0 ± 0.3	24.0 ± 3.0	140.3 ± 23.7	1970.1 ± 638.3	54.53	14.21^begkmp	2.10 ± 0.98^cekmp	57.35
	5	7/7	22.3 ± 0.5	22.5 ± 1.6	146.2 ± 26.5	1150.0 ± 510.6	31.69	8.26^cfiloq	1.39 ± 0.47^cfhlnr	71.80
	10	7/7	22.3 ± 0.5	23.7 ± 0.7	150.9 ± 32.6	1084.2 ± 502.8	28.37	7.39 ^cfiloq	0.90 ± 0.43^cfilor	81.67
	20	7/7	22.2 ± 0.3	22.3 ± 1.7	158.9 ± 22.6	664.3 ± 190.7	16.53	4.31^cfilor	0.73 ± 0.28 ^cfilor	85.20

vs Control, ^ap < .05; ^bp < .01; ^c p < .001；vs Vehicle, ^dp < .05; ^ep < .01; ^f p < .001；vs HyperCVAD + RTX, ^gp < .05; ^hp < .01; ⁱp < .001; vs LBH589, ^jp < .05; ^kp < .01; ^lp < .001; vs Hyper-CVAD, ^mp < .05; ⁿp < .01; ^op < .001; vs RTX ^pp < .05; ^qp < .01; ^rp < .001.

Table 5. Effect of PM for Injection on median survival of subcutaneously transplanted mice with human Burkitt's lymphoma Raji cell line.

Group	Control	Vehicle	PM 10mg/kg	PM 20mg/kg	HyperCVAD
Median survival (day)	14	14	30^bei	30 ^bei	16

vs Control, ^ap < .05; ^bp < .01; ^c p < .001; vs Vehicle, ^dp < .05; ^ep < .01; ^fp < .001; vs Hyper-CVAD, ^gp < .05; ^hp < .01; ⁱp < .001.

Table 6. Pharmacokinetic parameters of oral administration and intravenous administration PM solution at a dose of 10 mg/kg.

	p.o.	i.v.
Cmax (μg/L)	49.3 ± 26.443	3137 ± 1323
Tmax (min)	5.67 ± 1.97	4.67 ± 1.63
AUC (μg/L∗ h)	56.2 ± 24.57	837 ± 277
t1/2 (h)	1.62 ± 0.691	0.340 ± 0.328
CL (L/h/Kg)	437 ± 197.44	13.5 ± 6.10

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary materials.