Steven Weitman

Biodegradable block copolymers for delivery of proteins and water-insoluble drugs

Release of several drugs from new ABA-type biodegradable thermal gels, ReGel, including proteins and conventional molecules, are presented. These are biodegradable, biocompatible polymers that demonstrate reverse thermal gelation properties. Organic solvents are not used in the synthesis, purification, or formulation of these polymers. The unique characteristics of ReGel hinge on the following two key properties: (1) ReGel is a water soluble, biodegradable polymer at temperatures below the gel transition temperature; (2) ReGel forms a water-insoluble gel once injected. This is consistent with a hydrophobically bonded gel state where all interactions are physical, with no covalent crosslinking. An increase in viscosity of approximately 4 orders of magnitude accompanies the sol--gel transition. The gel forms a controlled release drug depot with delivery times ranging from 1 to 6 weeks. ReGels inherent ability to solubilize (400 to >2000-fold) and stabilize poorly soluble and sensitive drugs, including proteins is a substantial benefit. The gel provided excellent control of the release of paclitaxel for approximately 50 days. Direct intratumoral injection of ReGel/paclitaxel (OncoGel) results in a slow clearance of paclitaxel from the injection site with minimal distribution into any organ. Efficacies equivalent to maximum tolerated systemic dosing were observed at OncoGel doses that were 10-fold lower. Data on protein release (pGH, G-CSF, insulin, rHbsAg) and polymer biocompatibility are discussed.

Antitumor Activity of a Kinesin Inhibitor

Several members of the kinesin family of microtubule motor proteins play essential roles in mitotic spindle function and are potential targets for the discovery of novel antimitotic cancer therapies. KSP, also known as HsEg5, is a kinesin that plays an essential role in formation of a bipolar mitotic spindle and is required for cell cycle progression through mitosis. We identified a potent inhibitor of KSP, CK0106023, which causes mitotic arrest and growth inhibition in several human tumor cell lines. Here we show that CK0106023 is an allosteric inhibitor of KSP motor domain ATPase with a Ki of 12 nM. Among five kinesins tested, CK0106023 was specific for KSP. In tumor-bearing mice, CK0106023 exhibited antitumor activity comparable to or exceeding that of paclitaxel and caused the formation of monopolar mitotic figures identical to those produced in cultured cells. KSP was most abundant in proliferating human tissues and was absent from cultured postmitotic neurons. These findings are the first to demonstrate the feasibility of targeting mitotic kinesins for the treatment of cancer.

Discovery and development of clofarabine: a nucleoside analogue for treating cancer

The treatment of acute leukaemias, which are the most common paediatric cancers, has improved considerably in recent decades, with complete response rates approaching ∼90% in some cases. However, there remains a major need for treatments for patients who do not achieve or maintain complete remission, for whom the prognosis is very poor. In this article, we describe the challenges involved in the discovery and development of clofarabine, a second-generation nucleoside analogue that received accelerated approval from the US FDA at the end of 2004 for the treatment of paediatric patients 1–21 years old with relapsed or refractory acute lymphoblastic leukaemia after at least two prior regimens. It is the first such drug to be approved for paediatric leukaemia in more than a decade, and the first to receive approval for paediatric use before adult use.

Single-cell analysis of circulating tumor cells identifies cumulative expression patterns of EMT-related genes in metastatic prostate cancer

Prostate tumors shed circulating tumor cells (CTCs) into the blood stream. Increased evidence shows that CTCs are often present in metastatic prostate cancer and can be alternative sources for disease profiling and prognostication. Here we postulate that CTCs expressing genes related to epithelial–mesenchymal transition (EMT) are strong predictors of metastatic prostate cancer.

Direct Raman imaging techniques for study of the subcellular distribution of a drug

Direct Raman imaging techniques are demonstrated to study the drug distribution in living cells. The advantage of Raman imaging is that no external markers are required, which simplifies the sample preparation and minimally disturbs the drug mechanism during imaging. The major challenge in Raman imaging is the weak Raman signal. In this study, we present a Raman image model to describe the degradation of Raman signals by imaging processes. Using this model, we demonstrate special-purpose image-processing algorithms to restore the Raman images. The processing techniques are then applied to visualize the anticancer agent paclitaxel in living MDA-435 breast cancer cells. Raman images were obtained from a cancer cell before, during, and after drug treatment. The paclitaxel distribution illustrated in these images is explained by means of the binding characteristics of the paclitaxel and its molecular target-the microtubules. This result demonstrates that direct Raman imaging is a promising tool to study the distribution of a drug in living cells.

Effect of Nanonization on Absorption of 301029: Ex Vivo and In Vivo Pharmacokinetic Correlations Determined by Liquid Chromatography/Mass Spectrometry

AbstractPurpose. To compare Caco-2 monolayer permeability and in vivo bioavailability of microparticle with nanoparticle 301029, a thiadiazole derivative, and to determine whether nanonization could improve oral bioavailability of the poorly soluble compound. Methods. The mean particle size of 301029 was reduced from 7 μm to 280 nm by pearl milling. In the ex vivo assay, both microparticle and nanoparticle 301029 at the same concentration were separately added to apical side and were collected from basolateral side of Caco-2 monolayer. In the bioavailability study, the two particle sizes of 301029 were orally administered to rats, respectively, and blood samples were collected. Nanoparticle 301029 in culture medium and rat serum was detected by a liquid chomatography-mass spectrometer (LC/MS) coupled with atmospheric pressure chemical ionization (APCI). Results. Permeability rate and permeated amounts of nanoparticle 301029 across the Caco-2 monolayer were about four times higher than those of microparticle 301029. In a pharmacokinetic study, nanoparticle 301029 showed Tmax about 1 h, whereas the microparticle 301029 showed Tmax at 4 h. The Cmax and AUC of nanoparticle 301029 were 3- to 4-fold greater than those of microparticle 301029, resulting in a significant increase in oral bioavailability of 301029 as compared with microparticle 301029. The ex vivo permeability and in vivo pharmacokinetic data indicate that nanoparticle formulation improves both absorption rate and absorption extent of the poorly soluble drug. Conclusions. Nanoparticle formulation enhances both Caco-2 monolayer permeability and rat oral bioavailability of the poorly soluble 301029. The result also demonstrates a close correlation between ex vivo Caco-2 permeability model and in vivo gastrointestinal absorption.

Phase I Trial of Paclitaxel in Children With Refractory Solid Tumors: A Pediatric Oncology Group Study

PURPOSE A phase I study was performed to describe the principal toxicities and identify the maximum-tolerated dose (MTD) of Taxol (paclitaxel; Bristol-Myers Squibb Co, Wallingford, CT) in children with therapy-resistant solid tumors. Additionally, the pharmacokinetic disposition of Taxol in children was studied, and preliminary evidence of the activity of Taxol against pediatric solid tumors was assessed. PATIENTS AND METHODS Twenty-four-hour continuous infusions of Taxol were administered every 21 days to children (median age, 12 years; range, 2 to 22) with refractory solid tumors. Doses ranged from 200 to 420 mg/m2, there was no intrapatient dose escalation. RESULTS A total of 62 courses of Taxol were administered to 31 patients. Two patients developed acute anaphylaxis during their second infusion of taxol at doses of 200 mg/m2 and 350 mg/m2, respectively. No other allergic reactions were documented. Myelosuppression occurred at all dose levels, but was of short duration (< or = 7 days) and did not appear to increase with consecutive courses or at higher dosage levels. A stocking-and-glove peripheral neuropathy became evident at doses > or = 290 mg/m2. Dose-limiting neurotoxicity occurred at 420 mg/m2 and comprised a significant fine-motor and peripheral neuropathy in one patient, and a tonic-clonic seizure in another. End-of-infusion plasma concentrations ranged from 0.40 to 6.4 mumol/L, and were not found to be dose-dependent over the range of doses studied. A complete response was documented in one patient, partial response in two, and minimal response in one for an overall response rate of 13%. CONCLUSION Neurotoxicity was dose-limiting when Taxol was administered by 24-hour continuous infusion to pediatric patients with relapsed solid tumors. In this population, the recommended dose for phase II trials is 350 mg/m2/d.

Hepatocellular carcinoma in children associated with Gardner syndrome or familial adenomatous polyposis.

Purpose Gardner syndrome, a variant of familial adenomatous polyposis, is characterized by colonic polyps that undergo malignant change and benign and malignant extracolonic lesions. Tumors frequently associated with Gardner syndrome include carcinoma of the ampulla of Vater, papillary carcinoma of the thyroid, and, in children, hepatoblastoma. The childhood malignancies often precede the appearance of other manifestations by several years. Patients and Methods Two patients are described. Gardner syndrome was diagnosed in a 15-year-old girl with fibrolamellar hepatocellular carcinoma after desmoid tumors and colonic polyposis developed. Classic hepatocellular carcinoma was also diagnosed in a 9 1/2-year-old boy with familial adenomatous polyposis. Results In patient I, the diagnosis of fibrolamellar hepatocellular carcinoma preceded the diagnosis of Gardner syndrome by almost 2 years. The diagnosis was confirmed by identifying a germline mutation of the adenomatous polyposis coli (APC) gene. This is the first patient reported with fibrolamellar hepatocellular carcinoma associated with Gardner syndrome. Patient 2 had a strong family history of familial adenomatous polyposis but no manifestations of Gardner syndrome. He was not tested for the APC mutation. The current literature and previously reported cases of hepatocellular carcinoma in patients with Gardner syndrome or familial adenomatous polyposis are reviewed. Conclusions Because hepatocellular carcinoma is uncommon in the pediatric and adolescent population, it is important to consider the possibility of Gardner syndrome or familial adenomatous polyposis in these patients.

Phase I Therapy Trials in Children With Cancer

Purpose: This study examined the response and toxicity rates of antineoplastic drugs evaluated in phase I clinical trials in children to identify trends in response and toxicity over time. Patients and Methods: Full length, peer-reviewed articles describing the results of single agent phase I therapy trials in children younger than 21 years with cancer were reviewed. Tumor-specific response data and doses of drugs that resulted in objective responses were noted. Deaths that occurred on study caused by drug toxicity. progressive disease (PD). or complications of marrow aplasia were identified, along with drug doses that resulted in toxic death. Temporal trends in response rates, toxicity. and number of patients entered in trials were examined. Results: A total of 1.606 patients with cancer were enrolled in 56 single-agent pediatric phase I therapy trials published between 1978 and 1996. Of these. 1,257 were evaluated for response by tumor type. The overall objective response rate was 7.9%. Response rates were highest for patients with neuroblastoma (17.7%) and acute myelogenous leukemia (11.6%). Patients with osteosarcoma and rhabdomyosarcoma had response rates of <3%. Sixty percent of responses in patients with solid tumors occurred at 81 to 100% of the maximum tolerated dose (MTD). although 42% of responses in patients with leukemia occurred at >100% of the MTD. Death on study was noted in 7.0% of all patients entered in trials. Only 0.7% of patients experienced a death related to drug toxicity. PD accounted for the death of 5.6% of study participants. A trend of increasing response rate despite smaller trial size was noted over the last 7 years of this period. Conclusion: Phase I trials in children with cancer represent a safe mechanism to determine the MTD. toxicity profile, and pharmacokinetics of new agents for use in children with cancer.

The folate receptor as a potential therapeutic anticancer target

The folate receptor, a 38,000 Dalton glycosylphosphatidylinositol-linked membrane protein, has been identified qualitatively in selected normal tissues and is relatively overexpressed in certain neoplastic tissues [1]. Three receptor subtypes have been identified in humans: the high-affinity (Kd = nM) α-subtype which is found in the kidney and is relatively overexpressed in epithelial tumors, the more abundant, low-affinity (Kd = mM) β-subtype which is found in the placenta, and the γ -subtype which is the secretory form. The mechanism of folic acid uptake by the receptor, through a process termed potocytosis, has been well described [2]. While the role of the receptor in folate homeostasis has not been well delineated, it is thought to provide a selective advantage for tumor growth over normal tissue or, because of the glycosylphosphatidylinositol tail, it may have a role in cell activation or communication. Since the folate receptor is relatively overexpressed in epithelial tumors, particularly in ovarian tumors in adults, in ependymoma in children, and choroid plexus tumors in both populations, there has been considerable interest in exploiting the folate receptor in tumor imaging, drug and antisense delivery, immunotherapy, and antifolate drug design. It has been shown that folic acid, when covalently linked to other macromolecules, retains its high affinity for the folate receptor [3]. Radionuclides, antineoplastic agents, antibodies, antisense DNA and toxins have been targeted to cell lines and tumors which overexpress the folate receptor using folic acid conjugates. The purified receptor has been used to produce monospecific antibodies against choriocarcinoma, renal, lung and ovarian tumors [4,5,6] and bispecific antibody against ovarian tumors [7] which have shown potential in clinical trials as immunotherapy and radiotherapy. Recent studies of new antifolate compounds have evaluated their affinity for the folate receptor or the reduced folate carrier as well as for enzymes of the folate pathway [8,9]. The purpose of this paper is to review the current preclinical and clinical studies and to identify future directions for investigations of the receptor with respect to treatment of pediatric and adult tumors.