Efficacy, Pharmacokinetics, Biodistribution and Excretion of a Novel Acylated Long-Acting Insulin Analogue INS061 in Rats

Figures & data

Figure 1 Structure of INS061.

Table 1 AUC for Glucose at Different Time Periods After Subcutaneous Dosing of Insulins at a Dose of 100 Nmol/Kg in STZ-Induced Diabetic Rats (n=6)

AUC for Glucose (mg·h/dL)	Vehicle	INS061	Humulin R	Degludec	INS061 vs^a
					Vehicle	Humulin R	Degludec
AUC0-24h	11,754 ± 353.4	7369 ± 278.3	10,990 ± 415.5	7994 ± 533.7	< 0.01	< 0.01	< 0.05
AUC0-16h	7953 ± 202.4	4417 ± 201.6	7146 ± 321.1	4613 ± 303.5	< 0.01	< 0.01	> 0.05
AUC0-12h	5978 ± 138.3	3298 ± 157.5	5070 ± 259.4	3226 ± 214.8	< 0.01	< 0.01	> 0.05
AUC0-8h	3933 ± 90.8	2355 ± 115.3	3092 ± 174.8	2198 ± 164.5	< 0.01	< 0.01	> 0.05
AUC0-6h	2975 ± 78.4	1912 ± 98.2	2156 ± 132.0	1754 ± 148.4	< 0.01	< 0.05	> 0.05
AUC0-4h	2022 ± 41.6	1436 ± 69.2	1328 ± 100.1	1309 ± 100.8	< 0.01	< 0.05	< 0.05
AUC0-2h	995 ± 26.5	825 ± 33.0	724 ± 52.2	778 ± 43.2	< 0.01	< 0.05	> 0.05
AUC0-1h	476 ± 21	440 ± 20.6	421 ± 20.4	417 ± 20.2	< 0.05	> 0.05	> 0.05

Note:^aA two-side t test method was used for the statistical analysis.

Figure 2 Blood glucose-lowering profiles of insulins at a dose of 100 nmol/kg in STZ-induced diabetic rats. (A) Blood glucose-time profiles, (B) AUC for glucose-time profiles, ***p<0.01, ^##p<0.05. The values are expressed as an average with standard deviation.

Table 2 The Mean Non-Compartmental Pharmacokinetic Parameters of ¹²⁵I-INS061 After a Single Intravenous or Subcutaneous Injection in Rats by RA Method (n=6)

Parameter	Unit	Subcutaneous Injection			Intravenous Injection
		0.75 IU/kg	1.5 IU/kg	3.0 IU/kg	1.5 IU/kg
t1/2	h	6.06 ± 0.902	6.53 ± 1.64	6.38 ± 1.85	2.54±0.422
Tmax	h	3.50 ± 0.548	3.00 ± 0.632	2.33 ± 0.816	0.083
Cmax	ng/mL	30.09 ± 5.47	67.44 ± 9.13	137.12 ± 18.50	390.78±81.12
AUC0-t	ng/mL/h	237.43 ± 32.93	459.72 ± 65.73	880.08 ± 95.30	649.84±80.65
AUC0-∞	ng/mL/h	251.48 ± 35.82	485.26 ± 67.23	917.66 ± 95.39	656.29±77.05
MRT	h	8.14 ± 0.706	7.39 ± 0.745	6.56 ± 0.869	2.88±0.556
Vd (Vd/F)	L/kg	0.277 ± 0.122	0.261 ± 0.066	0.267 ± 0.069	0.076±0.024
Cl (Cl/F)	L/kg/h	0.031 ± 0.012	0.028 ± 0.004	0.029 ± 0.003	0.021±0.003
Fabs	%	-	73.94	-	-

Table 3 The Mean Non-Compartmental Pharmacokinetic Parameters of ¹²⁵I-INS061 After a Single Intravenous or Subcutaneous Injection in Rats by TCA-RA Method (n=6)

Parameter	Unit	Subcutaneous Injection			Intravenous Injection
		0.75 IU/kg	1.5 IU/kg	3.0 IU/kg	1.5 IU/kg
t1/2	h	5.61 ± 1.19	5.17 ± 1.05	4.81 ± 1.11	2.40 ± 0.61
Tmax	h	2.50 ± 0.548	2.33 ± 0.516	2.17 ± 0.753	0.083
Cmax	ng/mL	24.94 ± 6.06	54.61± 9.39	108.78 ± 20.36	339.73 ± 72.33
AUC0-t	ng/mL/h	146.72 ± 20.29	309.76 ± 50.49	617.74 ± 81.78	442.23±76.74
AUC0-∞	ng/mL/h	156.63 ± 22.51	323.31 ± 52.47	633.28 ± 82.87	446.75±77.10
MRT	h	7.64 ± 0.899	6.19 ± 0.181	5.6 ± 0.406	2.08±0.310
Vd (Vd/F)	L/kg	0.346 ± 0.070	0.308 ± 0.044	0.296 ± 0.075	0.103±0.020
Cl (Cl/F)	L/kg/h	0.043 ± 0.006	0.042 ± 0.006	0.043 ± 0.005	0.030±0.004
Fabs	%	-	70.04	-	-

Figure 3 Mean plasma concentration-time curves of INS061 after a single subcutaneous administration at the doses of 0.75, 1.5 and 3.0 IU/kg in rats (n=6). (A) The radioactivity was determined by RA method. (B) The radioactivity was determined by TCA-RA method. (C) Comparison of mean plasma concentration-time curves determined by RA, TCA-RA and HPLC-RA methods at 1.5 IU/kg dose in rats.

Table 4 Comparison of Main Pharmacokinetic Parameters Determined by RA, TCA-RA and HPLC-RA Methods After a Single Subcutaneous Injection of ¹²⁵I-INS061 at a Dose of 1.5 IU/Kg in Rats (n=6)

Parameter	Unit	HPLC-RA	TCA-RA	RA
t1/2	h	4.21 ± 1.69	5.17 ± 1.05	6.53 ± 1.64
Tmax	h	2.33 ± 0.516	2.33 ± 0.516	3.00 ± 0.632
Cmax	ng/mL	60.78 ± 7.50	54.61± 9.39	67.44 ± 9.13
AUC0-t	ng/mL/h	289.33 ± 34.39	309.76 ± 50.49	459.72 ± 65.73
AUC0-∞	ng/mL/h	303.06 ± 33.08	323.31 ± 52.47	485.26 ± 67.23
Cl (Cl/F)	L/kg/h	0.044 ± 0.005	0.042 ± 0.006	0.028 ± 0.004

Table 5 Comparison of the Tissue Distribution Determined by RA and TCA-RA Methods After Subcutaneous Injection of ¹²⁵I-INS061 in Rats

Tissue	AUC(0–24h) (ng/g/h)		Ratio (%)
	RA	TCA-RA
Heart	227.76	155.06	68.08
Lung	334.95	226.93	67.75
Liver	366.23	227.85	62.22
Spleen	208.57	152.11	72.93
Kidney	1054.65	482.32	45.73
Stomach	137.11	90.03	65.66
Intestine	215.16	159.79	74.27
Adipose	86.43	60.83	70.38
Muscle	293.56	172.28	58.69
Brain	38.65	25.88	66.96
Serum	496.43	394.32	79.43

Figure 4 Tissue distribution of total radioactivity at 0.5, 2, 8, and 24 h after a single subcutaneous administration at 1.5 IU/kg in rats (n=6). (A) The radioactivity was determined by RA method. (B) The radioactivity was determined by TCA-RA method.

Figure 5 Cumulative excretion after subcutaneous administration of ¹²⁵I-INS061 in rats. (A) urine and feces. (B) bile.