Pawel Robak

Current and emerging treatments for chronic lymphocytic leukaemia.

Chronic lymphocytic leukaemia (CLL) is the most common adult leukaemia in Europe and North America. The disease is characterized by proliferation and accumulation of small CD5+ B cells in blood, lymph nodes, spleen, liver and bone marrow. The natural clinical course of CLL is highly variable, and chemotherapy is usually not indicated in early and stable disease. However, patients with progressive and more advanced CLL require treatment. For many years, chlorambucil with or without corticosteroids was used in previously untreated patients with CLL. More recently, purine nucleoside analogues (PNAs) [fludarabine, cladribine and pentostatin] have been included in treatment approaches for this disease, and chlorambucil is no longer the leading standard everywhere. Currently, this drug is rather recommended for the treatment of older, unfit patients with co-morbidities, especially in European countries. Significantly higher overall response (OR) and complete response (CR) rates in patients treated initially with PNAs than in those treated with chlorambucil or cyclophosphamide-based combination regimens have been confirmed in randomized, prospective, multicentre trials. Moreover, PNAs administered in combination with cyclophosphamide produce higher response rates, including CR and molecular CR, compared with PNA as monotherapy.Recent reports suggest that the administration of monoclonal antibodies (mAbs) can significantly improve the course of CLL. At present, two mAbs have the most important clinical value in patients with CLL. The first is rituximab, a human mouse antibody that targets CD20 antigens, and the second is alemtuzumab, a humanized form of a rat antibody active against CD52. Several recent reports suggest that in patients with CLL, rituximab combined with a PNA can increase the OR and CR rates compared with PNA or rituximab alone, with acceptable toxicity. In randomized trials, the combination of rituximab with fludarabine and cyclophosphamide (FC-R regimen) demonstrated higher rates of OR, CR and progression-free survival in patients with previously untreated and relapsed or refractory CLL than fludarabine plus cyclophosphamide (FC regimen). Several reports have confirmed significant activity with alemtuzumab in relapsed or refractory CLL, as well as in previously untreated patients.Recently, several new agents have been investigated and have shown promise in treating patients with CLL. These treatments include new mAbs, agents targeting the antiapoptotic bcl-2 family of proteins and receptors involved in mediating survival signals from the microenvironment, antisense oligonucleotides and other agents. The most promising are new mAbs directed against the CD20 molecule, lumiliximab and anti-CD40 mAbs. Oblimersen, alvocidib (flavopiridol) and lenalidomide are also being evaluated both in preclinical studies and in early clinical trials. In recent years, a significant improvement in haematopoietic stem cell transplantation (HSCT) procedures in patients with high-risk CLL has been observed. However, the exact role of HSCT, autologous or allogeneic, in the standard management of CLL patients is still undefined.

Current Status of Older and New Purine Nucleoside Analogues in the Treatment of Lymphoproliferative Diseases

For the past few years more and more new cytotoxic agents active in the treatment of hematological malignancies have been synthesized and become available for either in vitro studies or clinical trials. Among them the class of antineoplastic drugs belonging to the purine nucleoside analogues group (PNAs) plays an important role. Three of them: pentostatin (DCF), cladribine (2-CdA) and fludarabine (FA) were approved by Food and Drug Administration (FDA) for the treatment of hematological malignancies. Recently three novel PNAs: clofarabine (CAFdA), nelarabine (ara-G) and forodesine (immucillin H, BCX-1777) have been synthesized and introduced into preclinical studies and clinical trials. These agents seem to be useful mainly for the treatment of human T-cell proliferative disorders and they are currently undergoing clinical trials in lymphoid malignancies. However, there are also several studies suggesting the role of these drugs in B-cell malignancies. This review will summarize current knowledge concerning the mechanism of action, pharmacologic properties, clinical activity and toxicity of PNAs accepted for use in clinical practice, as well as new agents available for clinical trials.

Older and new purine nucleoside analogs for patients with acute leukemias

Purine nucleoside analogs (PNAs) compose a class of cytotoxic drugs that have played an important role in the treatment of hematological neoplasms, especially lymphoid and myeloid malignancies. All PNA drugs have a chemical structure similar to adenosine or guanosine, and they have similar mechanisms of action. They have many intracellular targets: they act as antimetabolites, competing with natural nucleosides during DNA or RNA synthesis, and as inhibitors of key cell enzymes. In contrast to other antineoplastic drugs, PNAs act cytotoxically, both in the mitotic and quiescent cell cycle phases. In the last few years, three PNAs have been approved for the treatment of lymphoid malignancies and other hematological disorders: 2-chlorodeoxyadenosine (2-CdA), fludarabine and pentostatin. 2-CdA and fludarabine are also active in the treatment of acute myeloid leukemia (AML). These drugs, in combination with cytarabine and other agents, are commonly used as salvage regimens in relapsed or refractory AML. Moreover, the addition of 2-CdA to the standard induction regimen is associated with an increased rate of complete remission and improved survival of adult patients with AML. More recently three novel PNAs have been synthesized and introduced into clinical trials: clofarabine, nelarabine and forodesine. Clofarabine is the most promising PNA in current clinical trials in pediatric and adult patients with acute leukemias. Nelarabine is more cytotoxic in T-lineage than in B-lineage leukemias. Clofarabine and nelarabine have been approved for the treatment of refractory patients with acute lymphoblastic leukemia (ALL) and lymphoblastic lymphoma. Clofarabine is also an active drug in AML treatment when administered either alone or in combination regimens as front-line treatment and in relapsed or refractory patients. Unlike other PNA, forodesine is not incorporated into DNA but displays a highly selective purine nucleoside phosphorylase inhibitory action. Forodesine is undergoing clinical trials for the treatment of T-cell malignancies, including T-cell ALL. This article summarizes recent achievements in the mechanism of action, pharmacological properties and clinical activity and toxicity of PNAs, as well as their emerging role in lymphoid and myeloid acute leukemias.

BCR Signaling in Chronic Lymphocytic Leukemia and Related Inhibitors Currently in Clinical Studies

Normal B lymphocytes receive signals from B-cell antigen receptor (BCR) that are triggered by binding of the BCR to an external antigen. Tonic signaling through the BCR provides growth and signals to chronic lymphocytic leukemia (CLL) cells, and plays an important role in the pathogenesis and progression of the disease. Antigen engagement of BCR is followed by intracellular recruitment and activation of BCR-associated kinases including spleen tyrosine kinase (Syk), Brutons tyrosine kinase (Btk) and phosphatidylinositol 3-kinases (PI3K). Inhibition of signaling pathways downstream of the BCR induces disruption of chemokine-mediated CLL cell migration and cell killing. BCR signal transduction inhibitors represent a promising new strategy for targeted CLL treatment. A number of therapeutic agents have recently been developed with significant activity in CLL. The compounds that are currently investigated in patients with CLL include ibrutinib –inhibitor of Btk, fostamatinib-inhibitor of Syk and idelalisib (GS-1101) –a specific isoform of the PI3K (PI3K) inhibitor. The clinical activity of ibrutinib, GS-1101 and fostamatinib in patients with CLL is associated with marked lymphocytosis due to release of tumor cells from the lymph nodes into the peripheral blood. Further studies are ongoing with single agents and their combinations with other targeted and conventional therapies. This article will review the preclinical rationale of BCR signaling inhibitors in the treatment of CLL, and the clinical evidence supporting the use of these agents in CLL patients.

The role of non-steroidal anti-inflammatory drugs in the risk of development and treatment of hematologic malignancies

Non-steroidal anti-inflammatory drugs (NSAIDs) comprise the group of structurally diverse but similarly acting compounds that are used for relieving signs and symptoms of inflammation, especially in treatment of rheumatic diseases. Recent reports suggested potential association between regular use of NSAIDs and the risk of development of hematological malignancies. However, the data distinctly differ depending on type of NSAID used, period of its administration and type of malignancy. Regular use of aspirin and other NSAIDs was shown to correlate with reduced risk of lymphoid malignancies. Frequent use of aspirin was found to be associated with decreased risk of acute leukemia (AL) development. In contrast, correlation between long-term acetaminophen usage and increased incidence of AL and multiple myeloma (MM) was indicated. On the other hand, NSAIDs were found to exert anti-cancer effects, inhibiting proliferation and invasive growth or inducing cell apoptosis in several tumors, including hematologic malignancies. One of those agents, non-cyclooxygenase 2-inhibiting R-enantiomer of etodolac (SDX-101), exerts cytotoxic effects against chronic lymphocytic leukemia (CLL) and MM cells, and is currently investigated in phase II clinical trial in CLL. The indole-pyran analogue of SDX-101, SDX-308 (CEP-18082), showed more potent cytotoxicity than SDX-101 against MM cells and inhibited osteoclast formation and activity of mature osteoclasts. Thus, SDX-308 may be an ideal agent for bone disease in MM and related diseases. Another analogue of SDX-101, SDX-309, showed also significant anti-tumor activity in first preclinical studies. The potential role of NSAIDs in prevention and treatment of hematologic malignancies is the subject of this review.

Anti-CD37 antibodies for chronic lymphocytic leukemia

Introduction: Immunotherapy using mAbs is a safe and effective method for the treatment of chronic lymphocytic leukemia (CLL) and other lymphoid malignancies. In recent years, mAbs based on selective B-cell depletion – rituximab, ofatumumab and obinutuzumab – have been approved for use in CLL therapy. More recently, CD37, a member of the tetraspanin superfamily of cell surface antigens, has been considered as a target for B-cell malignancies. Areas covered: The results of preclinical and early clinical studies suggest that in patients with CLL, newer anti-CD37 agents, otlertuzumab (formerly known as TRU-016), BI 836826, IMGN529 and 177Lu-tetulomab can be useful in the treatment of this disease. Expert opinion: CD37 may offer advantages over CD20 as a target for CLL cells. It is selectively expressed on normal mature B cells and by most B-cell malignancies. Anti-CD37 antibodies may be useful for patients resistant or refractory to anti-CD20 mAb therapy or relapsing after such treatment. The development of these agents into a clinically useful therapy for CLL is probably many years away and will be followed with great interest by laboratory investigators and clinicians.

Subcutaneous versus intravenous bortezomib in patients with relapsed multiple myeloma: Subanalysis of patients with renal impairment in the phase III MMY-3021 study

Renal impairment is a common presenting feature in multiple myeloma (MM) and an estimated 50% of patients are affected during the course of their disease.[1][1] Moderate and/or severe renal impairment/failure is associated with poorer survival.[2][2]–[4][3] Subanalyses of phase III studies in

Rituximab plus fludarabine and cyclophosphamide or other agents in chronic lymphocytic leukemia.

Over the last few years, several monoclonal antibodies have been investigated in patients with B-cell lymphoid malignancies. Rituximab is the most important monoclonal antibody of clinical value in these disorders. Rituximab is an IgG1 chimeric antibody containing murine light- and heavy-chain variable region sequences and human constant region sequences. Since approval in 1997, rituximab has become the standard of care in follicular B-cell lymphoma, chronic lymphocytic leukemia (CLL) and aggressive lymphoma when combined with chemotherapy. Higher clinical benefits of rituximab can be seen in patients with CLL when it is added to other chemotherapeutic agents. Several recent reports have suggested that in patients with CLL, rituximab combined with purine nucleoside analogs (PNAs) or PNAs and cyclophosphamide may improve the results with acceptable toxicity, both in previously untreated and refractory/relapsed patients. The randomized, multinational Phase III study (REACH trial) has shown that rituximab combined with fludarabine and cyclophosphamide (R-FC regimen) results in 10 months longer progression-free survival, and higher overall response and complete response rates than fludarabine and cyclophosphamide (FC regimen) in previously treated patients. The German CLL study group initiated a multicenter, multinational Phase III trial, CLL8, to evaluate the efficacy and tolerability of R-FC versus FC for the first-line treatment of patients with advanced CLL. The overall response rate was significantly higher in the R-FC arm (95%) compared with FC (88%). The complete response rate in the R-FC arm was 44% compared with 27% in the FC arm. The recently updated analysis has demonstrated longer overall survival in the R-FC group. Recent clinical observations have revealed that combinations of rituximab with pentostatin and cyclophosphamide, or cladribine and cyclophosphamide are also highly active regimens in previously untreated CLL. In addition, the results of treatment with high-dose methylprednisolone in combination with rituximab in advanced CLL resistant to fludarabine have been reported recently by several groups. However, available therapies are only partially effective in CLL, exposing an obvious need to develop new, more specific and active drugs. Recently, several new anti-CD20 monoclonal antibodies have been developed and are now being evaluated in clinical trials.

Antibody therapy alone and in combination with targeted drugs in chronic lymphocytic leukemia

The development of non-chemotherapeutic agents, including monoclonal antibodies (mAbs) and other targeted drugs, makes chemotherapy-free treatment an attractive option for chronic lymphocytic leukemia (CLL). The classical mAb, rituximab, has been authorized for use in both first-line and second-line therapy for CLL. New mAbs directed against CD20, ofatumumab, and obinutuzumab (GA-101) have also been approved for the treatment of this disease. Recently, several new mAbs with potential benefits over the approved anti-CD20 antibodies have been developed for use in CLL. Anti-CD37, anti-CD19, and anti-CD40 mAbs are in early clinical trials and show promise in treating CLL. In addition, the combination of mAbs with B-cell receptor signaling pathway inhibitors and immunomodulatory drugs makes the chemotherapy-free option a reality today. Combinations of antibodies with targeted drugs like ibrutinib, idelalisib, or lenalidomide are expected to replace chemotherapy-based combinations for treating CLL in the near future. However, phase III trials should confirm the benefit of these new treatment strategies and establish their exact place in the therapeutic armamentarium for CLL.

High activity of rituximab combined with cladribine and cyclophosphamide in a patient with pulmonary lymphomatoid granulomatosis and bone marrow involvement.

Lymphomatoid granulomatosis (LG) is a rare T-cell rich, B-cell angiodestructive non-Hodgkin’s lymphoma that involves mainly the lungs, skin, liver, kidneys and the nervous system [1]. The prognosis is generally poor and median survival is *4 years [2]. The therapeutic approach and optimal management have not been well defined. Various approaches have been reported including steroids alone or combined with cyclophosphamide and combination chemotherapy [2]. Other treatment options include interferon-a (IFN-a), cyclosporin A and bone marrow transplantation [3 – 5]. More recently, a few cases treated with anti-CD20 monoclonal antibody rituximab have been reported with good response in some of them [6 – 10]. We present a patient who developed pulmonary LG with bone marrow involvement and only partially responded to IFN-a therapy. Subsequently, she was successfully treated with rituximab combined with cladribine (2-CdA) and cyclophosphamide (RCC). The patient was a 51-year-old white female, first seen in our outpatient clinic in March 2004 because of chronic non-productive cough, fever, fatigue, over 10% body weight loss and asthenia in the last 8 months. The main finding on physical examination was splenomegaly and hepatomegaly 1 cm below the costal margin. The complete blood count revealed hemoglobin 9.5 g dl, red blood cells 4.06 10 l, hematocrite 30.7%, white blood cells 8.56 10 l and platelets 2676 10 l. The bone marrow aspirate was hypercellular. Clusters of mature and partially immature lymphoid cells with a prominent nucleolus were observed (Figure 1(a)). In the bone marrow trephine biopsy there were nodular infiltration of large B-cells which were CD20þ, and some of them CD30 positive. The erythrocyte sedimentation rate was 42 after 1 h and C reactive protein was 96.5 mg l (normal values 0 – 5 mg). The b2-microglobulin level was increased (6110 ng ml) and lactate dehydrogenase activity was also increased (341 U L). The chest X-ray showed irregular infiltrations in lower lobes of both lungs. Computed tomography of the chest also showed multiple nodules ranging from 5 – 10 mm in diameter to larger infiltrates and atelectatic changes in lower lobes of both lungs. An open lung biopsy was performed and two pieces of lung tissue were examined. Histologically, these nodules contained lymphoid tissue consisting of the mixture of small and larger lymphocytes and admixed histiocytes and neutrophiles (Figure 1(b)). Larger cells presented B-cell antigens (CD20, CD79a) and a proportion of them were also CD30positive. There were also foci of neutrophiles, mostly located close to blood vessels. The wall of blood vessels was infiltrated with B-cells but fibrinoid necrosis was not observed. Smaller cells were of T origin (CD3-positive). Polymerase chain reaction