Activity In Several Malignancies Research Articles

Asiaticoside Inhibits Growth and Metastasis in Non-Small Cell Lung Cancer by Disrupting EMT via Wnt/β-Catenin Pathway.

Non-small cell lung cancer (NSCLC) is the primary inducer of cancer-related death worldwide. Asiaticoside (ATS) is a triterpenoid saponin that has been indicated to possess an antitumor activity in several malignancies. Nonetheless, its detailed functions in NSCLC remain unclarified. In this study, NSCLC cells were exposed to various doses of ATS. Functional experiments were employed to estimate the ATS effect on NSCLC cell behaviors. Western blotting was implemented for protein expression evaluation. A xenograft mouse model was established to assess the ATS effect on NSCLC in vivo. The results showed that ATS restrained NSCLC cell proliferation, cell cycle progression, migration, and invasiveness. ATS reversed TGF-β-induced promotion in epithelial-mesenchymal transition (EMT). Mechanistically, ATS inhibited Wnt/β-catenin signaling in NSCLC. Upregulating β-catenin restored ATS-mediated suppression of NSCLC cell aggressiveness. Moreover, ATS administration repressed tumorigenesis in tumor-bearing mice. In conclusion, ATS represses growth and metastasis in NSCLC by blocking EMT via the inhibition of Wnt/β-catenin signaling.

Constitutive type-1 interferons signaling activity in malignant gliomas.

Recent studies revealed a pro-tumor effect of constitutive Type-1 interferons (IFN-I) production and the downstream signaling activity in several malignancies. In contrast, heterogeneity and clinical significance of the signaling activity in gliomas remain unknown. Thus, we aimed to depict the heterogeneity and clinical significance of constitutive Type-1 interferon (IFN-I) production and the downstream signaling activity in gliomas. We utilized multiplex immunofluorescence (mIF) on a 364 gliomas tissue microarray from our cohort. Moreover, we conducted bioinformatic analyses on the Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA) databases to investigate the heterogeneity and clinical significance of constitutive IFN-I signaling activity in gliomas. We observed high heterogeneity of the constitutive IFN-I signaling activity among glioma subtypes. Signaling increased with the WHO malignancy grade while decreasing in the gliomas with IDH mutations. Additionally, high IFN-I activity served as an independent predictor of unfavorable outcomes, and global DNA hypermethylation in IDH-mutant gliomas was associated with decreased IFN-I signaling activity. Positive correlations were observed between the IFN-I activity and glioma-associated inflammation, encompassing both anti-tumor and pro-tumor immune responses. Furthermore, the IFN-I activity varied significantly among tumor and immune cells in the glioma microenvironment (GME). Notably, a distinct pattern of IFN-I signaling activity distribution in GME cells was observed among glioma subtypes, and the pattern was independently associated with patient overall survival. Constitutive IFN-I signaling activity varies significantly among glioma subtypes and represents a potential indicator for increased glioma inflammation and unfavorable clinical outcomes.

The Potent Dihydroorotate Dehydrogenase Inhibitor, Hosu-53, Exhibits Compelling Monotherapy Efficacy in Multiple Myeloma and Augments CD47 Targeted Therapy

Worldwide, multiple myeloma (MM) is the second most common hematological malignancy characterized by the expansion of aberrant mature plasma cells in the bone marrow. Despite the FDA approval of several drugs with different mechanisms of action such as immunomodulatory drugs (IMiDs) and proteosome inhibitors (PIs), MM remains a challenging incurable disease where most patients ultimately relapse and become refractory to available . The resistant nature of MM underscores the need for novel therapeutic strategies that can directly affect MM cells while enhancing the outcome with MM current standard of care (SOC). Among the most promising MM SOC agents is monoclonal antibodies (mAb) such as the CD38 antibody daratumumab, which significantly improved the management of newly diagnosed and relapsed/refractory MM. Therefore, we aimed to develop novel therapies that can enhance mAb therapy in MM. Dihydroorotate dehydrogenase (DHODH) is an enzyme which mediates the fourth and rate-limiting step in the de novo pyrimidine synthesis pathway converting dihydroorotate to orotate, the precursor of uridine. Pyrimidine starvation using DHODH inhibitors has been shown to have potent antiproliferative activity in several malignancies including acute myeloid leukemia (AML) and MM. Thus, we developed a novel potent DHODH inhibitor, HOSU-53 for the treatment of hematological malignancies. We previously discovered the ability of HOSU-53 to modulate surface CD38 expression and found that HOSU-53 had potent monotherapy activity in MM and provided impressive synergy in combination with daratumumab using the NCI-H929 MM cell line derived xenograft (CDX) model (2022 AACR annual meeting abstracts). Herein, we further expanded our studies to pre-clinically develop HOSU-53 as a new potential MM therapy and explore additional synergistic immunotherapy combinations that could maintain efficacy in the case of anti-CD38 therapy resistance. We evaluated the in vitro antiproliferative efficacy of HOSU-53 against a panel of MM cell lines and found nanomolar potency validating that HOSU-53 would have monotherapy efficacy. Indeed, we conducted in vivo studies using two additional MM CDX subcutaneous (s.c) models, OPM-2 and RPMI-8226, and found significant tumor delay and survival advantage. In the OPM-2 model, HOSU-53 has a median survival of 54-days compared to vehicle at 28-days, while the median survival for HOSU-53 was 60-days compared to vehicle at 26-days in the RPMI-8226 model. We further verified HOSU-53 efficacy using the disseminated MM1.S luciferase CDX model and found a significant prolonged survival in the HOSU-53 cohort (median survival53-days) compared to vehicle (median survival28-days) that was further enhanced with isatuximab combination resulting in superior survival benefit (median survival 69-days). Given the continuous challenge in maintaining a durable response with anti-CD38 therapies due to resistance, we sought to explore additional combination regimens to maximize HOSU-53 efficacy. Currently there is significant clinical interest in CD47 antibody therapy such as magrolimab for both solid tumors and hematological malignancies. Our previous work suggested a strong synergy between CD47 antibodies and HOSU-53 in AML with curative potential (2022 ASH annual meeting abstracts), and hypothesized that we would observe similar synergy with HOSU-53 in combination with anti-CD47 in MM. Indeed, we found complete tumor regression in all mice treated with HOSU-53 + B6.H12 anti-CD47 therapy in the NCI-H929 s.c CDX model and significant prolonged survival and reduced bioluminescence in the NCI-H929 luciferase disseminated CDX model. Together, these two studies validate a potential clinical benefit to explore HOSU-53 + anti-CD47 regimen in MM patients. Furthermore, we observed that calreticulin (pro-phagocytosis signal) was modulated post HOSU-53 in vitro treatment suggesting its role in the observed synergy between HOSU-53 and CD47 blockade therapy. In summary, we show compelling survival benefit for HOSU-53 as a monotherapy which is further enhanced when combined with anti-CD38 or anti-CD47 therapies. HOSU-53 is expected to enter phase 1 clinical trials in 2024 and our data is supportive for its expansion into MM.

Harmine suppresses the malignant phenotypes and PI3K activity in breast cancer

Breast cancer remains a serious threaten to the women's health, discovery of potent treatment would help to improve the outcomes of breast cancer patients. Harmine extracted from Peganum harmala L , has been reported to exert tumor suppressive activity in several malignancies. Our objective was to demonstrate the effects of harmine on the malignant phenotypes of breast cancer cells. Breast cancer cell lines (MDA-MB-231, SKBR3, and MCF-7) and human normal breast cell line MCF-10A were employed in the present study. The MTT and colony formation assays were applied to the detection of cell viability and proliferation. Wound healing and transwell assays were performed to evaluate the alterations of cell migration and invasion after harmine treatment. Flow cytometry was applied to assess the effect of harmine in inducing cell apoptosis. Furthermore, western blotting assay was used to detect the biomarkers of epithelial-mesenchymal transition and phosphatidylinositol 3 kinase (PI3K) signaling pathway. The tumorigenesis ability was detected by subcutaneous implantation. Harmine dose-dependently suppressed the viability and proliferative capacity of breast cancer cells. Flow cytometry showed that harmine induced apoptosis in MCF-7 and MDA-MB-231 cells. In addition, harmine effectively inhibited the migration and invasion abilities of breast cancer cells. Western blotting indicated harmine significantly promoted E-cadherin and PTEN expression, while suppressed N-cadherin, vimentin, PI3K, p-mTOR, and AKT levels. Interfering the PTEN expression by siRNA partly rescued the activity of PI3K signaling pathway. Moreover, harmine injection also suppressed the tumorigenesis of breast cancer cells. Our results suggested that Hermine could suppress multiple malignant phenotypes and inhibit PI3K signaling, which supports that harmine might be a potential tumor-suppressive natural compound against breast cancer.

Optimization of Voyager V1 (VV1) oncolytic virus systemic delivery in combination with cemiplimab and ipilimumab in patients with melanoma and non–small cell lung cancer (NSCLC).

TPS9595 Background: There is a need for novel immunotherapies to address the patient population that never or no longer responds to immune checkpoint inhibitors (CPI). VV1 is an oncolytic vesicular stomatitis virus engineered to express human interferon beta (IFNβ) to enhance cellular anti-tumor immune responses and tumor selectivity. Phase 1 studies demonstrated VV1 anti-tumor activity in several malignancies with or without a CPI. We are exploring ways to optimize VV1 efficacy in combination with cemiplimab, an anti-PD1 antibody approved for lung, basal and squamous cell skin cancers. Recent clinical data support a 5-fold higher dose of VV1 than was previously explored, and pre-clinical data show synergy between the oncolytic and an anti-CTLA4 antibody, in addition to cemiplimab, supporting exploration of a triplet. What was originally a five-arm study of intravenous (IV) VV1 in combination with IV cemiplimab has been amended to focus on 2 means of optimizing efficacy: use of a higher dose of VV1 and triplet combination in proof-of-concept populations. Methods: We are now enrolling pts with advanced melanoma (after progression on anti-PD1) and plan to include 1st-line NSCLC pts with PD-L1 expression ≥50%. The study is first exploring the preliminary anti-tumor activity, safety, and immunogenic activity of the combination of IV VV1 at a dose of 1.0 x 1011 TCID50 once on D1 followed by IV cemiplimab Q3W starting on D8, or the same regimen with an additional intratumoral injection of VV1 1.0 x 109 TCID50 once on D1 for pts with accessible lesions. Pts receive IV cemiplimab Q3W until confirmed disease progression or intolerable toxicity. Once at least 6 pts have been treated with acceptable safety across the 2 melanoma doublet cohorts using this higher dose of VV1, a 3rd melanoma cohort will open to add a single dose of ipilimumab 50 mg on D1 (all IV triplet). Once 6 melanoma pts have received the triplet safely, the 1st-line NSCLC cohort will open. All cohort decisions are guided by a Data Review Committee. Cohorts will be expanded based on a Simon 2-stage design using a type I error rate of 0.05 and power of 85%. Null ORR is 10% with a target of 35% for 2nd line melanoma and null ORR is 40% in 1st line NSCLC with a target of 70%. Each melanoma cohort will require a response in ≥2 of 10 pts in the 1st stage to add 11 more in the 2nd stage, while NSCLC will first need 5/9 evaluable pts to respond, then an additional 13 to complete the design. Response is assessed at week 7 then Q9W per RECIST v1.1. The study includes serial biopsies in ≥3/10 pts in Stage 1 of each of the IV melanoma cohorts (doublet and triplet therapy), all pts in Stage 2 of these IV melanoma cohorts, and all pts in both Stage 1 and Stage 2 of the IV/IT melanoma cohort, to permit a thorough investigation of the impact of the 3 immunotherapies under investigation. The study is currently ongoing in the USA. Clinical trial information: NCT04291105.

Cyclin-dependent Kinases 4/6 Inhibitors in Neuroendocrine Neoplasms: from Bench to Bedside.

Cyclin-dependent kinases (CDKs) are key regulators that play an important role in cell division. Palbociclib, ribociclib and abemaciclib showed significant antitumor activity in several malignancies and, recently, also a myeloprotective effect for trilaciclib when added to chemotherapy. The purpose of this review is to highlight the current evidence for CDK4/6 inhibitors in neuroendocrine neoplasms (NENs). Preclinical results showed a promising antitumor activity of CDK4/6 inhibitors in neuroendocrine tumors (NETs), but so far, the very few small clinical trials did not show a strong impact on progression free survival (PFS) and objective response in NETs. Meanwhile, the CDK4/6 inhibitor trilaciclib revealed significant effects in reducing chemotherapy-induced myelosuppression in small cell lung cancer (SCLC). Up to date, CDK4/6 inhibitors are still considered investigational in NETs as antitumor agents, whereas trilaciclib can be used in the routine clinical practice in extensive stage SCLC patients for reducing myelotoxicity of standard chemotherapy.

Pretreatment prognostic nutritional index is a prognostic marker for extensive-stage small cell lung cancer patients treated with anlotinib.

BackgroundAnlotinib is a multi-targeted tyrosine kinase inhibitor that inhibits tumor angiogenesis which has shown activity in several malignancies and approved for the treatment of small cell lung cancer (SCLC) in China. However, there are no markers can predict the clinical outcomes of anlotinib. We aimed to evaluate the efficacy of anlotinib in extensive stage SCLC (ES-SCLC) patients who failed at least two regimens treatment and to explore potential factors related to its survival benefit.MethodsPatients with ES-SCLC treated with anlotinib monotherapy were screened between March 2017 and May 2019, prognostic nutritional index (PNI) before treatment were collected. Progression free survival (PFS) and overall survival (OS) were calculated and compared using the Kaplan–Meier method and the log-rank test. The prognostic values of each variable were evaluated with univariate and multivariate Cox proportional hazard regression (PHR) analyses.ResultsA total of 41 patients with ES-SCLC were received anlotinib treatment, the median age was 57 (range, 33–76). Median OS was significantly longer in the PNI high arm compared with the low arm [8.4 months (95% CI, 5.1–9.6 months) vs. 4.7 months (95% CI, 2.1–6.3 months); hazard ratio (HR) 0.42 (95% CI, 0.21–0.85); P=0.01]. The median PFS of two arms were 4.1 months (95% CI, 2.1–8.2 months) and 2.6 months (95% CI, 0.7–3.9 months), respectively (HR =0.53, 95% CI, 0.27–1.02, P=0.05). Multivariate analysis confirmed that PNI (P<0.01) and LDH (P<0.01) were significant independent biomarkers for OS.ConclusionsThe present study demonstrated that pretreatment PNI can be used as a novel and convenient biomarker to predict the prognosis in ES-SCLC patients treated with anlotinib.

PF366 CXCR5+PD‐1+ CD8+ T‐CELLS IN HUMAN BLOOD AND LYMPH NODES

Background:Re‐invigorating exhausted T‐cells through immune checkpoint blockade (ICB) has shown promising activity in several malignancies. However, response rates vary between malignancies, and it is unclear which parameters predict response to ICB. Recently, a subset of CXCR5+PD‐1+ CD8+ T‐cells has been identified in mice. This memory‐like population appears to be crucial for the expansion of CD8+ T‐cells during ICB, and is characterized by expression of the transcription factor Tcf1. CXCR5+ CD8+ T‐cells have also been identified in humans, but it's currently unknown whether this distinctive CD8+ T cell subset has a similar function as described in mice, and plays an equally important role during ICB.Aims:To investigate the human CXCR5+PD‐1+ CD8+ T‐cell subset, we studied peripheral blood and lymph node samples derived from healthy donors and patients with a variety of B cell malignancies.Methods:PB and LN samples were analyzed by flow cytometry.Results:The frequency of CXCR5+PD‐1+ CD8+ T‐cells was higher in lymph nodes. CXCR5+PD‐1+ CD8+ T‐cells resemble antigen‐experienced memory‐like cells, with high expression of KLRG1, CD127, granzyme K, and Eomes, and low expression of effector molecules granzyme B and T‐bet. They express high levels of Tcf1, in accordance with the phenotype described in mice. CXCR5+PD‐1+ CD8+ T‐cells were present within the EBV‐specific T‐cell population, especially directed against epitopes expressed during viral latency. In line with mice data, we could not detect CXCR5+PD‐1+ CD8+ T‐cells within the FLU‐specific T‐cell compartment, suggesting that chronic antigen stimulation is required for this population to develop. On a functional level, CXCR5+PD‐1+ CD8+ T cells produce effector cytokines IL‐2, IFNγ and TNFα after stimulation, in line with their memory‐like phenotype. CXCR5+PD‐1+ CD8+ T cells proliferate and differentiate into effector CD8+ T cells, with higher expression of granzyme B and loss of Tcf1.We assessed the role of CXCR5+PD‐1+ CD8+ T cells in ICB by analyzing their phenotype in samples derived from patients with various B cell malignancies with differential clinical response rates to ICB. In chronic lymphocytic leukemia (CLL), a disease with no objective response to ICB, CXCR5+PD‐1+ CD8+ T cells express lower levels of Tcf1 and markers compatible with an advanced differentiation state, and express lower levels of PD‐1. We did not observe these changes in CXCR5+PD‐1+ CD8+ T cells in B cell malignancies with better response rates to ICB, like DLBCL, multiple myeloma, and follicular lymphoma. Currently, we perform functional experiments in the context of checkpoint blockade to verify whether this changed phenotype is related to the disappointing response towards ICB in CLL. To explore the potential role of CXCR5+PD‐1+ CD8+ T cells for ICB therapy further, we are analyzing peripheral blood samples derived from cancer patients who receive ICB therapy while also studying the localization and frequency of CXCR5+PD‐1+ CD8+ T‐cells in relation to tumor cells within a spectrum of malignant tissues by confocal microscopy.Summary/Conclusion:In conclusion, we find that on a phenotypical and functional level, human CXCR5+PD‐1+ CD8+ T‐cells closely resemble this subset in mice. In CLL, in which ICB therapy does not lead to clinical responses, CXCR5+PD‐1+ CD8+ T‐cells have an altered phenotype which might explain the lack of response to ICB. Furthermore, if human CXCR5+PD‐1+ CD8+ T‐cells indeed play a similar important role in the response to ICB therapy as in mice, they could potentially serve as a biomarker for clinical response in human malignancies.

Ramucirumab: A vascular endothelial growth factor receptor-2 inhibitor with activity in several malignancies.

The pharmacology, pharmacokinetics, safety, dosage and administration, current and potential roles in therapy, and cost considerations of ramucirumab are reviewed. Ramucirumab is a recombinant monoclonal antibody that selectively inhibits vascular endothelial growth factor receptor-2. Ramucirumab has been approved for the treatment of gastric cancer, non-small-cell lung cancer (NSCLC), and metastatic colorectal cancer. Ramucirumab displays similar pharmacokinetics in patients with gastric cancer, NSCLC, and metastatic colorectal cancer. The most common adverse effects of any grade with ramucirumab monotherapy included fatigue, decreased appetite, abdominal pain, hypertension, anemia, hemorrhage, and diarrhea. For ramucirumab monotherapy in patients with gastric cancer, ramucirumab 8 mg/kg should be administered by i.v. infusion every two weeks. For combination therapy, ramucirumab should be administered at the same dose with weekly paclitaxel 80 mg/m(2) i.v. on days 1, 8, and 15 every 28 days. When treating NSCLC, the recommended ramucirumab dose is 10 mg/kg i.v. with docetaxel 75 mg/m(2) i.v. on day 1 every 21 days. For combination therapy with FOLFIRI (fluorouracil, leucovorin, and irinotecan) for metastatic colorectal cancer, ramucirumab 8 mg/kg should be administered by i.v. infusion every two weeks. The average wholesale prices of ramucirumab are $1224 and $6120 for the 100- and 500-mg single-dose vials, respectively. Ramucirumab has demonstrated benefit in a variety of cancers, supporting the role of antiangiogenic agents in the management of malignancies. However, the clinical benefit in certain malignancies may be offset by the high cost of ramucirumab and the duration of treatment.

An open‐label, single‐arm, phase 2 trial of the polo‐like kinase inhibitor volasertib (BI 6727) in patients with locally advanced or metastatic urothelial cancer

Polo-like kinases (Plks) control multiple steps during the cell cycle, and Plk1 is overexpressed in urothelial cancer (UC). Volasertib (BI 6727), a Plk inhibitor, has demonstrated antitumor activity in several malignancies, including UC. In this phase 2 trial, the authors investigated volasertib as a second-line treatment in advanced/metastatic UC. Patients who progressed within 2 years of 1 prior chemotherapy regimen received 300 mg volasertib on day 1 every 3 weeks. The dose was escalated to 350 mg in cycle 2 if volasertib was tolerated in cycle 1. The primary endpoint was tumor response, which was assessed every 6 weeks; secondary endpoints were progression-free survival, overall survival, duration of response, safety, and pharmacokinetics. Fifty patients were enrolled, and the median patient age was 68.5 years (range, 52-83 years). All patients had received prior platinum, 94% of patients had relapsed ≤2 years after prior therapy, 36% had liver metastases, and 54% had lung metastases. The median number of treatment cycles was 2 (range, 1-27 treatment cycles), and 23 patients were dose escalated at cycle 2. Seven patients (14%) had a partial response, 13 (26%) had stable disease, and 30 (60%) progressed within 6 weeks. The median response duration was 41 weeks (range, 29.1-77.3 weeks). The median progression-free survival was 1.4 months, and the median overall survival was 8.5 months. The most frequent grade 3 and 4 adverse events were neutropenia (28%), thrombocytopenia (20%), and anemia (16%). No cumulative toxicity was observed. Volasertib as second-line treatment for advanced/metastatic UC had an acceptable safety profile but demonstrated insufficient antitumor activity for further evaluation as a monotherapy.

Silvestrol exhibits significant in vivo and in vitro antileukemic activities and inhibits FLT3 and miR-155 expressions in acute myeloid leukemia

BackgroundActivating mutations [internal tandem duplication (ITD)] or overexpression of the FMS-like tyrosine kinase receptor-3 (FLT3) gene are associated with poor outcome in acute myeloid leukemia (AML) patients, underscoring the need for novel therapeutic approaches. The natural product silvestrol has potent antitumor activity in several malignancies, but its therapeutic impact on distinct molecular high-risk AML subsets remains to be fully investigated. We examined here the preclinical activity of silvestrol in FLT3-ITD and FLT3 wild-type (wt) AML.MethodsSilvestrol in vitro anti-leukemic activity was examined by colorimetric cell viability assay, colony-forming and flow cytometry assays assessing growth inhibition and apoptosis, respectively. Pharmacological activity of silvestrol on FLT3 mRNA translation, mRNA and protein expression was determined by RNA-immunoprecipitation, qRT-PCR and immunoblot analyses, respectively. Silvestrol in vivo efficacy was investigated using MV4-11 leukemia-engrafted mice.ResultsSilvestrol shows antileukemia activity at nanomolar concentrations both in FLT3-wt overexpressing (THP-1) and FLT3-ITD (MV4-11) expressing AML cell lines (IC50 = 3.8 and 2.7 nM, respectively) and patients’ primary blasts [IC50 = ~12 nM (FLT3-wt) and ~5 nM (FLT3-ITD)]. Silvestrol increased apoptosis (~4fold, P = 0.0001), and inhibited colony-formation (100%, P < 0.0001) in primary blasts. Silvestrol efficiently inhibited FLT3 translation reducing FLT3 protein expression by 80–90% and decreased miR-155 levels (~60%), a frequently co-regulated onco-miR in FLT3-ITD-positive AML. The median survival of silvestrol-treated vs vehicle-treated mice was 63 vs 29 days post-engraftment, respectively (P < 0.0001).ConclusionsSilvestrol exhibits significant in vivo and in vitro antileukemic activities in AML through a novel mechanism resulting in inhibition of FLT3 and miR-155 expression. These encouraging results warrant a rapid translation of silvestrol for clinical testing in AML.

The immunological era in melanoma treatment: new challenges for heat shock protein-based vaccine in the advanced disease

Introduction: Tumor-derived heat shock protein (HSP)–peptide complexes (HSPPCs) induced immunity against malignancies in preclinical trials, working across tumor types and bypassing the need to identify single immunogenic peptides. These results paved the way for the use of human gp96 obtained from autologous tumor samples as an anti-cancer vaccine.Areas covered: Autologous tumor-derived HSP gp96 peptide complex (HSPPC-96) vaccine is emerging as a tumor- and patient-specific cancer vaccine, with confirmed activity in several malignancies. It has been tested in Phase III clinical trials in advanced melanoma and kidney cancer with evidence for efficacy in patients with earlier stage disease. HSPPC-96-based vaccine demonstrated an excellent safety profile, thus emerging as a novel therapeutic approach with a suggestive role in cancer therapy. This review summarizes work on the use of HSPPC-96 as an autologous anti-tumor vaccine in advanced melanoma. Data were retrieved by PubMed and Medline research and using the authors' personal experience.Expert opinion: Further investigations are needed to understand the biological basis of immune functions in order to improve the clinical outcome of HSP-based cancer therapy. In the near future, the combination of HSP-based vaccines with other biological compounds might represent a successful strategy in the therapy of advanced melanoma.

C-myb activates &amp;lt;italic&amp;gt;CXCL12&amp;lt;/italic&amp;gt; transcription in T47D and MCF7 breast cancer cells

Chemokine C-X-C motif ligand 12 (CXCL12) is a potent chemotactic and angiogenic factor that has been proposed to play a role in organ-specific metastasis and angiogenic activity in several malignancies. In this study, we found that the overexpression of c-myb could elevate CXCL12 mRNA level and CXCL12 promoter activity in human T47D and MCF-7 breast cancer cells. Chromatin immunoprecipitation assay demonstrated that c-myb could bind to the CXCL12 promoter in the cells transfected with cmyb expression vector. c-myb siRNA attenuated CXCL12 promoter activity and the binding of c-myb to the CXCL12 promoter in T47D and MCF-7 cells. These results indicated that c-myb could activate CXCL12 promoter transcription.

Phase I/II study of bortezomib (B) in patients with systemic AL-amyloidosis (AL)

8050 Background: AL is characterized by deposition of monoclonal immunoglobulin light chains leading to multi-organ failure and death. B (Velcade®) is a specific inhibitor of the 26S proteasome with activity in several malignancies. This mechanism of action raises concern of enhancing the amyloidogenic protein deposition and worsening of AL. This phase I/II study is to determine the maximum tolerated dose (MTD) and toxicity profile of B in pts previously treated for AL. Secondary objectives include the rate and duration of hematologic and organ responses. Methods: A 3+3 dose escalation design was used for the phase I part. Eligibility criteria included previously treated AL with organ involvement, LVEF =40% and adequate organ function. Pts with prior B treatment, infection, significant cardiac disease, bleeding or gr. 2 neuropathy were excluded. Seven cohorts are planned to examine two administration schedules. A weekly schedule administering 0.7,1.0,1.3 or 1.6 mg/m2 of B on day 1, 8, 15, 22 of a 35-day cycle. In the bi-weekly schedule, 0.7, 1.0 or 1.3 mg/m2 are given on day 1,4,8, 11 of a 21-day cycle. Pts were assessed at specified intervals and dose-limiting toxicities (DLT) were assessed during cycle 1. Results: At this time, 15 pts were treated in the 4 weekly cohorts. Median age was 59 yrs (38–74). All pts had previously received systemic therapy; 100% had received melphalan (58% with prior SCT), 83% steroids, 58% thalidomide. 75% of pts had = 2 organs involved at baseline, renal 42%, cardiac 33%, neuropathy 33%. One DLT (worsening CHF) was observed in cohort 1.6 mg/m2. Two pts discontinued early due to CHF and hypotension. Grade 3/4 AEs were reported in 42% of pts and the most frequent were fatigue 17%, infection 17%, CHF 8% and retinal detachment 8%. The most frequent grade 1/2 AEs were diarrhea 67%, nausea 42%, fever 33%, dizziness 33% and neuropathy 8%. There were no treatment-related deaths. Of the 11 evaluable pts for response, 5 (45%) had objective hematological responses (2 CR and 3 PR) and 4 pts (36%) have stable disease &gt; 6 months. Dose escalation on the biweekly schedule is ongoing. Conclusions: B has significant activity in patients with relapsed AL in this preliminary analysis and tolerance to B is satisfactory. The MTD for the weekly schedule is 1.6 mg/m2. Further assessment is planned in the phase II part. [Table: see text]

Phase I/II trial of bortezimib and pemetrexed in patients with advanced non-small cell lung cancer

18145 Background: Bortezomib (Vc) is a novel proteosome inhibitor with activity in several malignancies including multiple myeloma, mantle cell lymphoma, and NSCLC. In NSCLC, Vc has additive activity combined with carboplatin and gemcitabine in first line and with docetaxel in second line treatment. Pemetrexed (P) is active in NSCLC and preclinical data suggests a pro-apoptotic synergy between Vc and P. Therefore, we initiated a phase I/II dose finding study of Vc + P in previously-treated patients (pts) with advanced or metastatic NSCLC. Methods: Fifteen pts have been accrued to 3 of 4 planned dose levels of Vc + P. Starting doses (and # pts treated) were Vc 1.4 mg/m2 day 1 &amp; 8 + P 400 mg/m2 day 1 every 3 weeks (3 pts). The 2nd and 3rd dose levels were Vc 1.6 mg/m2 day 1 &amp; 8 + P 500 mg/m2 day 1 (8 pts, 5 new + 3 from dose level 1) and Vc 1.8 mg/m2 day 1 &amp; 8 + P 500 mg/m2 day 1 (7 pts). Results: 15 pts are evaluable for response and toxicity and include 8 males, 7 females, median age 67 (range, 55–82), PS 0/1 (3/12 pts), median of 2 prior therapies (range 1–3). Confirmed PRs occurred in 2 pts (13%) and stable disease in 5 (33%). Dose limiting toxicities consisted of grade 4 fatigue (1 pt) and neutropenia/fever (1 pt) at dose level 2, and grade 3 abdominal pain and fatigue (1 pt) and grade 3 diarrhea and vomiting (1 pt) at dose level 3. Conclusions: The above combination is safe at the doses tested thus far and active in pts with heavily pretreated, advanced NSCLC. We are currently exploring Vc 2.0 mg.m2 Day 1 &amp; 8 + P 500 mg/m2 day 1 every 3 weeks to determine the MTD and plan a multi-site Phase II study to determine response rate and survival in a larger pt population. No significant financial relationships to disclose.

Podophyllotoxin: Current Perspectives

Podophyllotoxin is a naturally occurring lignan with important antineoplastic and antiviral properties and supported by detailed understanding of their mechanism of action, and facilitated by chemical manipulations that have amplified their bioactivity, the podophyllotoxin analogues have advanced to the forefront of several areas of therapeutic and developmental chemotherapy. Additive and synergistic laboratory interactions with other cytotoxic drugs have been exploited to allow development of podophyllotoxin-based multidrug regimens, which are showing important activity in several malignancies, and many of its related analogues will complement conventional pharmaceuticals in treatment, prevention and diaganosis of disease, while at the same time adding value to agriculture. Additive and synergistic laboratory interactions with other cytotoxic drugs have been exploited to allow development of etoposide-based multidrug regimens, which are showing important activity in several malignancies. Extensive structural modifications of podophyllotoxin have been performed in order to obtain more potent and less toxic antitumour agents, which resulted in the widespread clinical introduction of two semisynthetic glucoconjugate analogues of etoposide and teniposide and newer agents with promising preclinical activity are in various stages of clinical assessment. As knowledge of molecular and biochemical mechanisms of action and resistance continues to expand, newer and better podophyllotoxin-based strategies for treatment of malignant disease are likely to evolve. This review provides a detailed discussion of research advances in the synthetic and medicinal chemistry of podophyllotoxin, and addresses the short history and pharmacological action of these compounds and further outlines the preclinical development and clinical trials of drugs in the pipeline and marketing approval. Finally, a systemic evaluation of novel and important analogues of podophyllotoxin and their contribution to the current structure-activity profile are considered. It is hoped that this review will be able to address the contributions of podophyllotoxin-related research to overall drug discovery and development and the role that this field will play in future. Keywords: Reviews, podophyllotoxin, short chronology, biological activity, medicinal application, mechanism of actions, chemical modification, structure-activity relationships.

Farnesyl transferase inhibitors: the next targeted therapies for breast cancer?

The ras family of proto-oncogenes are upstream mediators of several essential cellular signal transduction pathways involved in cell proliferation and survival. Point mutations of ras oncogenes result in constitutively active Ras and have been shown to be oncogenic. However, ras activation can occur in the absence of ras mutations secondary to upstream receptor activation. The first important step in Ras activation is farnesylation by farnesyl transferase, and inhibitors of this enzyme have been demonstrated to inhibit Ras signaling, and have anti-tumor effects. However, it is now clear that farnesyl transferase inhibitors (FTIs) have activity independent of Ras, most likely due to effects on prenylated proteins downstream of Ras, which explains their activity in several malignancies, including breast cancer, where ras mutations are rare. Several FTIs are in clinical development for the treatment of solid tumors. Preclinical evidence suggests that FTIs can inhibit breast cancers in vitro and in vivo, and a phase II trial of the FTI, R115777, in patients with advanced breast cancer produced encouraging results. Based on prior successful outcomes with agents targeting the estrogen and epidermal growth factor receptor pathways in breast cancer, the FTIs, used alone or more likely with other agents, may be the next exciting targeted therapy in breast cancer.

Phase I dose and sequencing study of pegylated liposomal doxorubicin and docetaxel in patients with advanced malignancies

Pegylated liposomal doxorubicin (PEG-LD) and docetaxel have single-agent activity in several malignancies. The authors conducted a Phase I trial to evaluate the maximum tolerated dose (MTD), toxicities, and effect of dose sequencing of this combination in patients with advanced malignancies. Twenty-two patients were enrolled in this two-arm, accelerated, dose escalation trial. Both drugs were administered on Days 1 and 15 of a 28 day cycle. In Arm A, dose escalation proceeded from a sequence and starting dose of 15 mg/m(2) PEG-LD and 30 mg/m(2) docetaxel. In Arm B, dose escalation proceeded from a sequence and starting dose of 30 mg/m(2) docetaxel and 15 mg/m(2)PEG-LD. In both arms, the dose of each drug was increased alternately by 5 mg/m(2) at each dose level. The MTD for Arm A was 20 mg/m(2) PEG-LD and 40 mg/m(2) docetaxel, both of which were administered on Days 1 and 15 of a 28-day cycle. The MTD for Arm B was 35 mg/m(2) docetaxel and 20 mg/m(2) PEG-LD, both of which were administered on Days 1 and 15 of a 28-day cycle. Dose-limiting toxicities were Grade 3 (according to the National Cancer Institute Common Toxicity Criteria) skin toxicity and thrombocytopenia. One partial response was observed and stable disease was documented for three patients. The recommended sequence and dose is 20 mg/m(2) PEG-LD followed by 40 mg/m(2) docetaxel on Days 1 and 15 of a 28-day cycle in Phase II trials for patients with breast and ovarian carcinoma to establish the efficacy of this well tolerated regimen.

A Phase II Trial of Gemcitabine in the Treatment of Advanced Bile Duct and Periampullary Carcinomas

Background: Gemcitabine is a novel nucleoside analogue with clinical anticancer activity in several malignancies. From September 1998 to April 2000, we treated patients with advanced bile duct and periampullary carcinomas with gemcitabine alone. Methods: Gemcitabine 1,000 mg/m²/day was administered in 200 ml of normal saline as a 30-min intravenous infusion on day 1 weekly for 3 weeks, followed by a 1-week rest. Results: A total of 24 consecutive patients (15 men, 9 women), with a median age of 59.5 years (range 40–72 years), were enrolled. All patients were evaluable for response: 1 patient achieved complete remission (CR); 2 patients had partial remission (PR); 8 patients remained stable (SD), and 13 patients had progressive disease (PD). The overall response rate (CR + PR) was 12.5% with a 95% confidence interval (CI) of 2.7–32.4%. The median progression-free survival (PFS) was 2.5 months (95% CI 1.6–5.5 months), and the median overall survival (OS) was 7.2 months (95% CI 3.8–8.9 months). Patients with disease control (CR + PR + SD) had better PFS and OS than those with PD. There were no treatment-related deaths. Few patients encountered grade 3/4 toxicity. Conclusion: Chemotherapy with gemcitabine demonstrated notable activity and was associated with a well-tolerable toxicity profile in patients with advanced biliary tract malignancies.

Acute oxaliplatin-induced peripheral nerve hyperexcitability.

Oxaliplatin is a novel platinum compound with clinical activity in several malignancies. Neurotoxicity is dose-limiting and occurs in two distinct forms, an acute neurologic symptom complex that occurs within hours or days of therapy and a chronic, cumulative sensory neuropathy. Patients were treated in a phase I study designed to establish the maximum-tolerated dose of capecitabine given with oxaliplatin. Because of the unusual neurosensory toxicity of oxaliplatin, detailed neurologic examination, needle electromyography (EMG), and nerve conduction studies (NCS) were performed before and the day after oxaliplatin in a subset of 13 patients. Carbamazepine therapy was tried in 12 additional patients to determine whether the neurologic effects might be relieved. All patients experienced acute, reversible neurotoxicities with oxaliplatin. Symptoms included paresthesias, dysesthesias, cold hypersensitivity, jaw pain, eye pain, pain in the arm used for drug infusion, ptosis, leg cramps, and visual and voice changes. Serial EMG and NCS revealed striking signs of hyperexcitability in motor nerves after oxaliplatin. In patients who achieved therapeutic levels, carbamazepine did not alter the clinical or electromyographic abnormalities. The acute neurotoxicity seen with oxaliplatin is characterized by peripheral-nerve hyperexcitability, and the findings are similar to the clinical manifestations of neuromyotonia. Carbamezepine, which provides symptomatic relief in acquired neuromytonia, did not seem to be beneficial. Efforts to identify a successful neuroprotectant strategy would have a major impact on improving patient quality of life and the ability to deliver full doses of oxaliplatin.