Preclinical Assessment of the Efficacy of Mycograb, a Human Recombinant Antibody against Fungal HSP90

Ruth C. Matthews; Gordon Rigg; Samantha Hodgetts; Tracey Carter; Caroline Chapman; Carl Gregory; Chris Illidge; James Burnie

doi:10.1128/AAC.47.7.2208-2216.2003

DOI: 10.1128/AAC.47.7.2208-2216.2003

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FIG. 1.
Lane 1, C. albicans extract immunoblotted against Mycograb; lane 2, molecular mass markers (31, 52, 98, and 185 kDa; Invitrogen, Paisley, United Kingdom); lane 3, silver stain of purified Mycograb showing monomer (28 kDa) and dimer (52 kDa).
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FIG. 2.
Two-dimensional electrophoresis and Western blot of C. albicans extract versus Mycograb.

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TABLE 1.

Checkerboard assay of FLC and Mycograb versus yeast isolates examined

Species	Agent	MIC (μg/ml) of each agent		FIC (μg/ml)	FICI	Outcome
Species	Agent	Alone	In combination	FIC (μg/ml)	FICI	Outcome
C. albicans (FLC-S)	FLC	1.56	0.4	0.26	0.34	Synergy
	Mycograb	128	10	0.08
C. albicans (FLC-R)	FLC	50	12.5	0.25	0.64	Indifference
	Mycograb	256	100	0.39
C. krusei	FLC	100	50	0.5	1.28	Indifference
	Mycograb	128	100	0.78
C. tropicalis	FLC	3.12	3.12	1.0	1.78	Indifference
	Mycograb	128	100	0.78
C. glabrata	FLC	6.25	1.56	0.25	0.75	Indifference
	Mycograb	128	64	0.5
C. parapsilosis (NCPF3104)	FLC	1.56	1.56	1	1.39	Indifference
	Mycograb	256	100	0.39
C. parapsilosis (clinical isolate)	FLC	1.56	1.56	1	1.39	Indifference
	Mycograb	256	100	0.39

TABLE 2.

Checkerboard assay of AMB and Mycograb versus yeast isolates examined

Species	Agent	MIC (μg/ml) of each agent		FIC (μg/ml)	FICI	Outcome
Species	Agent	Alone	In combination	FIC (μg/ml)	FICI	Outcome
C. albicans (FLC-S)	AMB	1	0.03	0.03	0.09	Synergy
	Mycograb	128	8	0.06
C. albicans (FLC-R)	AMB	0.5	0.125	0.25	0.27	Synergy
	Mycograb	256	4	0.02
C. krusei	AMB	1	0.25	0.25	0.28	Synergy
	Mycograb	128	4	0.03
C. tropicalis	AMB	0.125	0.016	0.13	0.16	Synergy
	Mycograb	128	4	0.03
C. glabrata	AMB	1	0.25	0.25	0.31	Synergy
	Mycograb	128	8	0.062
C. parapsilosis (NCPF3104)	AMB	1	0.25	0.25	0.27	Synergy
	Mycograb	256	4	0.016
C. parapsilosis (clinical isolate)	AMB	1	0.25	0.25	0.27	Synergy
	Mycograb	256	4	0.016

TABLE 3.

Pharmacokinetic parameters of Mycograb in mice given a single dose

Mycograb dose (mg/kg) (sample studied)	C_max (μg/ml)	AUC_0-t (μg · min/ml)	AUC_0-∞ (μg · min/ml)	t_1/2α (min)	t_1/2β (h)	MRT (h)
2 (serum)	4.7	155	157	3.75	2.34	3.17
10 (serum)	54.9	2,857.4	2,859.6	7.05	2.51	2.35
10 (renal biopsy specimen)	21.7	9,108	9,120	10.2	10.94	9.3

TABLE 4.

Organ colony counts from a series of experiments

Expt no.^b	Group	Dose (mg/kg)		Colony count (mean log₁₀ CFU/g ± SD)^a
Expt no.^b	Group	AMB	Mycograb	Kidney	Liver	Spleen
1	1	0.6	2	(9)^e	(9)^d	(9)^d
	2	0.6	0.2	(10)^e	(10)^d	(9)^d
	3	0.6		^b3.66 ± 0.82 (4)	(9)^d	(10)^d
	4		2	^b4.58 ± 0.76 (2)	^b3.50 ± 0.47 (3)	^b3.12 ± 0.11 (4)
	5		0.2	5.1 ± 0.11 (0)	3.85 ± 0.81 (3)	3.35 ± 0.33 (3)
	6			5.5 ± 0.5 (0)	4.42 ± 0.59 (2)	3.65 ± 0.69 (2)
2	1	0.6	2	(9)^d	(9)^d	(8)^d^,^e
	2	0.6	0.2	(9)^d	(8)^d	3.71 ± 0.64 (4)
	3	0.6		(9)^d	(7)	^b4.18 ± 0.22 (2)
	4		2	(6)	^b4.32 ± 0.82 (4)	^b3.87 ± 0.93 (1)
	5		0.2	(6)	^b4.35 ± 0.26 (4)	^b3.81 ± 0.78 (1)
	6			4.05 ± 0.1 (0)	5.15 ± 0.18 (0)	4.81 ± 0.33 (0)
3	1	0.6	2	(9)^d	(6)	(8)^d^,^e
	2	0.6	0.2	(10)^d	(6)	(8)^d^,^e
	3	0.6		(9)^d	^b4.74 ± 1.0 (2)	^b4.24 ± 0.25 (3)
	4		2	(8)^d	^b5.54 ± 0.18 (0)	^b5.19 ± 0.31 (0)
	5		0.2	5.17 ± 0.19 (2)	^b6.01 ± 0.25 (0)	5.30 ± 0.39 (0)
	6			5.19 ± 0.90 (0)	6.57 ± 0.34 (0)	5.86 ± 0.76 (0)
4	1	0.6	2	(8)^d	(10)^d	(10)^d
	2	0.6	0.2	(8)^d	(10)^d	(10)^d
	3	0.6		(8)^d	(10)^d	(10)^d
	4		2	^b4.24 ± 0.95 (1)	(9)^d	(10)^d
	5		0.2	^b4.41 ± 0.72 (1)	(10)^d	(10)^d
	6			5.30 ± 0.13 (0)	3.82 ± 0.69 (1)	4.41 ± 0.93 (3)
5	1	0.6	2	3.82 ± 0.81 (4)	(10)^d^,^e	(10)^d^,^e
	2	0.6	0.2	3.84 ± 0.78 (1)	(9)^d	3.89 ± 0.94 (2)
	3	0.6		4.01 ± 0.93 (1)	3.08 ± 0.12 (5)	4.10 ± 0.98 (0)
	4		2	4.01 ± 0.92 (1)	(9)^d	4.03 ± 0.33 (2)
	5		0.2	4.01 ± 0.09 (3)	(9)^d	4.07 ± 0.31 (3)
	6			4.00 ± 0.04 (0)	3.54 ± 0.65 (3)	4.05 ± 0.08 (0)
6	1	0.6	2	^b4.56 ± 0.81 (0)	^b3.66 ± 0.25 (4)	^b4.08 ± 0.02 (2)
	2	0.6	0.2	^b4.48 ± 0.34 (0)	^b4.09 ± 0.27 (2)	^b4.11 ± 0.29 (1)
	3	0.6		^b4.62 ± 0.94 (0)	^b3.98 ± 0.06 (3)	^b4.24 ± 0.29 (2)
	4		2	^b4.60 ± 0.65 (0)	^b4.07 ± 0.26 (3)	^b4.32 ± 0.31 (1)
	5		0.2	^b4.77 ± 0.79 (0)	^b4.28 ± 0.28 (0)	^b4.48 ± 0.38 (2)
	6			5.66 ± 0.64 (0)	5.31 ± 0.44 (0)	5.24 ± 0.16 (0)

↵a Number of negative biopsy specimen in parentheses.
↵b Experiments: 1, FLC-resistant C. albicans; 2, C. krusei; 3, C. glabrata 4, C. tropicalis; 5, C. parapsilosis; 6, C. parapsilosis (clinical isolate).
c Statistically significant (Scheffe's test, P < 0.05) versus the control group 6.
↵d Statistically significant number of negative biopsy specimens (Fisher's exact test, P < 0.05) versus the control group 6.
↵e Statistically significant numbers of negative biopsy specimens (Fisher's exact test, P < 0.05) versus the AMB-treated group 3.