SUPPLEMENTARY MATERIALS AND METHODS
Inclusion and exclusion criteria. The inclusion criteria determine patient eligibility for
the study were: a) histologic confirmation of recurrent anaplastic astrocytoma,
glioblastoma multiforme or gliosarcoma that is progressive despite previous radio- or
chemotherapy; b) brain tumor > 1.0 cm in diameter able to evaluated by gadoliniumenhanced MRI scan of the brain performed within 14 days of G207 administration and
the proposed area of initial study drug administration appeared to be surgically resectable
en bloc; c) candidate for brain tumor resection; d) failed external beam radiation therapy
≥ 5000 cGy at least 4 weeks prior to G207 administration; e) Karnofsky Performance
Status (KPS) ≥ 70%; f) age  18 years; g) stable steroid regimen within 1 week prior to
G207 administration; h) females with a negative urine pregnancy test within 24 hours
prior to G207 administration; i) willing to use effective barrier birth control to prevent
pregnancy and to guard against potential HSV viral shedding during the study and for a
minimum of six months following administration of G207; and, j) able and willing to
give written informed consent.
Subjects with any of the following conditions were ineligible: a) known HIV
seropositivity; b) documented extracranial metastases; c) multiple intracranial malignant
glioma lesions; d) tumor involvement which would have required ventricular, brainstem,
basal ganglia, or posterior fossa inoculation, or access through a ventricle to deliver study
drug; e) tumor position that could, in the Investigator’s opinion, have posed the risk of
penetration of the cerebral ventricular system during inoculation with study drug; f)
tumor involving both hemispheres or subependyma, or if CSF dissemination was
suspected; g) any contraindication for undergoing MRI such as: individuals with
pacemakers, epicardial pacer wires, infusion pumps, surgical and/or aneurysm clips,
shrapnel, metal prosthesis, implants with potential magnetic properties, metallic bodies in
the eyes (i.e., former welders), etc.; h) white blood cells (WBC) ≤ 3.0x103/mm3, absolute
neutrophil count (ANC) ≤ 1.5x103/mm3, platelets ≤ 100,000/mm3, hemoglobin ≤ 9.0
gm/dL, protime/international normalized ratio (INR) or partial thromboplastin time (PTT)
> 1.3 x control; i) serum creatinine > 1.7 mg/dL; j) liver transaminases aspartate
aminotransferase (AST) and/or alanine aminotransaminase (ALT) > 4x the upper limits
of normal, total and direct bilirubin > 1.5 mg/dL; k) prior brain tumor resection within 4
weeks prior to G207 administration; l) chemotherapy or cytotoxic therapy, or
immunotherapy (e.g., IL-2, IL-12, interferon) within 6 weeks prior to G207
administration; m) vaccinations within 30 days prior to G207 administration; n) history of
alcohol or other substance abuse; o) history of or current diagnosis of other cancer except
curative cervical cancer in-situ, basal or squamous cell carcinoma of the skin; p) history
of encephalitis, multiple sclerosis, other CNS infection or primary CNS disease that
would interfere with subject evaluation; q) history or current diagnosis of any medical or
psychological condition that in the Investigator's opinion, might interfere with the
subject’s ability to participate; r) history of prior gene transfer therapy or prior therapy
with cytolytic virus of any type; s) evidence of active herpes infection; t) current
treatment with any antiviral agent active against HSV (acyclovir, valacyclovir,
penciclovir, famciclovir, ganciclovir, foscarnet, cidofovir); u) female who was pregnant
and/or nursing; v) granulocytopenia, unstable and/or severe intercurrent medical
conditions or active uncontrolled infection that would have precluded surgery; w) history
of any investigational agent within 30 days prior to G207 administration. Two patients
did not meet the eligibility criteria but both received waivers. Subjects #101-107 and -108
met exclusion criterion No. 11 (chemotherapy or cytotoxic therapy within 6 weeks prior
to G207 administration).
Production of G207 using current Good Manufacturing Practices. Two different lots of
G207 were used in this study. These lots are thought to be bioequivalent after dilution to
the correct concentration for administration. In both virus lots, the host cells were derived
from a Vero Cell Bank, acquired from World Health Organization (WHO) Vero cells
(African green monkey kidney). For clinical Lot 2A, the upstream process was roller
bottle based and utilized a freeze-thaw step to release virus from cells. Major purification
and concentration steps in the downstream process were achieved using size exclusion
chromatography and ultracentrifugation, Clinical Lot 4 was produced in Vero cell culture
in a packed bed bioreactor, and released from cells with hypertonic (0.4 M sodium
chloride buffer). Subsequently, virus was purified and formulated by size exclusion
chromatography and ultrafiltration. Concentrated virus was filter sterilized and aliquoted.
The virus was then stored in 1.0 ml cryovials containing 0.12 ml of G207 suspended in
the storage buffer D-PBS/10% glycerin at -60C