FORMULATION AND EVALUATION OF DASATINIB TABLETS

FORMULATION AND EVALUATION OF DASATINIB TABLETS

AIM-To Formulate And Evaluate Dasatinib Tablets OBJECTIVE-To formulate and evaluate the dasatinib tablets to decrease side effects by
using various polymers for immediare release.

PLAN OF WORK Preformulation studies Compatibility studies Formulation development In-vitro evaluation tests

INTRODUCTION
Dasatinib, previously known as BMS-354825, is a cancer drug produced by Bristol-Myers Squibb and sold under the trade name Sprycel. Dasatinib is an oral multi- BCR/ABL and Src family tyrosine kinase inhibitor approved for use in patients with chronic myelogenous leukemia (CML) after imatinib treatment and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph+ ALL). It is being evaluated for use in numerous other cancers, including advanced prostate cancer. Chronic myelogenous leukemia (CML) is a chronic myeloproliferative disorder with an initially chronic course lasting for 3-5 years. It eventually transforms into accelerated and blastic phases, which are generally fatal. CML was one of the first diseases in which a specific chromosomal abnormality was identified, a t(9;22)(q34;q11) or Philadelphia chromosome. CML had been traditionally treated with conventional chemotherapy with hydroxyurea or busulfan. Although these agents can achieve hematologic remissions in most patients, no evidence of sustained disappearance of the chromosomal abnormality was evident. Interferon alpha (IFN-alpha) has been able to achieve hematologic and cytogenic remissions in a significant number of patients, and recent studies show a survival advantage for patients treated with IFN-alpha compared with those treated with conventional chemotherapy. The results of these studies are discussed, and the reasons for discordance among different investigators analyzed in this review. Allogeneic bone marrow

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FORMULATION AND EVALUATION OF DASATINIB TABLETS transplantation (BMT) may be curative in some patients with CML. The benefits and limitations of this approach in the treatment of CML are also discussed and the results of different alternatives compared. Other alternatives of therapy, including newer

chemotherapeutic agents, combinations of IFN-alpha with other agents, and autologous BMT, are presented. The availability of very sensitive techniques for detection of the Philadelphia chromosome at the molecular level has allowed the detection of minimal residual disease. The information available on these measurements is also analyzed. Finally, we discuss the alternatives for patients with accelerated and blastic phase CML, as well as the clinical characteristics and prognosis for patients with Philadelphia-chromosome-negative CML. Philadelphia chromosome (Ph)-positive acute lymphocytic leukemia (ALL) is characterized by a reciprocal translocation between the long arms of chromosomes 9 and 22 [t(9;22)(q34;q11)], leading to the formation of the Bcr-Abl fusion protein. This translocation occurs in about 3% to 5% of children and 20% to 30% of adults with ALL, and is associated with a very poor prognosis.1,2 Modern chemotherapy regimens for adult Ph-negative ALL result in complete response (CR) rates of 90% to 95% and long-term overall survival (OS) rates of 35% to 40%.3-7 Despite achievement of similarly high CR rates of 80% to 90% with these chemotherapy regimens, most patients with Ph-positive ALL relapse and die from the disease.8-10 The 5-year survival rates are less than 10% for adults. Children with Ph-positive ALL treated with chemotherapy alone have higher survival rates of 25% to 30%, but worse outcomes are seen with leukocyte counts higher than 100 109/L and/or failure to respond to glucocorticoid therapy.11,12 Allogeneic stem cell transplantation (SCT) in first CR can increase the long-term event-free survival (EFS) rate to 28% to 40% for adults and to 60% to 75% for children.11,13-18 The unique biologic behavior of Ph-positive ALL is attributed to molecular changes resulting from the Abl proto-oncogene from chromosome 9 fusing within the 5 half of the breakpoint cluster region (bcr) sequences on chromosome 22.19,20 The chimeric Bcr-Abl oncogene leads to fusion proteins of different sizes depending on the location of the Bcr breakpoint. If the break within the first intron of BCR (m-bcr region) splices the first exon of the BCR gene to the second exon of the ABL gene (e1a2), the 190 kDa fusion oncoprotein is generated. This oncoprotein (p190bcr-abl) occurs in 60% to 80% of patients with Ph-positive ALL; the other 20% to 40% are characterized by the larger p210bcr-abl fusion oncoprotein typical of chronic
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FORMULATION AND EVALUATION OF DASATINIB TABLETS myelogenous leukemia (CML). The latter is formed when exon b2 or exon b3 of the BCR gene (M-bcr region) is coupled to ABL exon 2 (b2a2 or b3a2 junction). Gleissner et al21 reported an association of the p210bcr-abl subtype with older age and trend toward worse 3year survival rates, although most studies have not demonstrated differences in clinical presentation or outcome with either p190bcr-abl or p210bcr-abl ALL.21-24 These Bcr-Abl molecular abnormalities may be sufficient for leukemic transformation as demonstrated in animal models showing a p210bcr-abl tyrosine kinase-dependent development of a CML-like disease.25 The mechanism of leukemogenesis appears related to the constitutively activated and dysregulated tyrosine kinase activity that results in downstream effects on multiple signal transduction pathways controlling cellular proliferation and apoptosis.26 Imatinib mesylate (Gleevec, STI571; Novartis Pharmaceuticals Corporation) is a selective inhibitor of the Bcr-Abl tyrosine kinase, platelet-derived growth factor (PDGF) receptor tyrosine kinase, and the c-kit receptor kinase.27 In vitro, imatinib mesylate inhibited the growth of Bcr-Abl-positive cells by inducing apoptosis and suppressing proliferation.28-30 In clinical trials, imatinib mesylate demonstrated significant anti-CML activity and is now a standard frontline treatment in newly diagnosed Ph-positive CML.31-39 Single-agent imatinib mesylate therapy for previously treated relapsed or refractory Ph-positive ALL and CML in lymphoid blast phase was associated with objective response rates of 40% to 50%; true CR rates (lasting for 4 weeks or more) were 5% to 7% with the median survival of 2 to 5 months.
39-41

Mechanisms of resistance such as Bcr-Abl amplification or mutations in the Bcr-Abl tyrosine kinase have been identified in previously treated Ph-positive ALL patients salvaged with single-agent imatinib mesylate.42,43 These results suggest that use of imatinib mesylate as a single agent in Ph-positive ALL is unlikely to be sufficient for long-term disease control. In vitro studies suggest synergistic activity of imatinib mesylate with chemotherapy agents such as anthracyclines, vincristine, and cytarabine (ara-C).44,45 This report summarizes the experience with hyper-CVAD (cyclophosphamide, vincristine, Adriamycin, and dexamethasone),46 a dose-intensive chemotherapy regimen used at our institution for adult ALL since 1992, and imatinib mesylate for patients with de novo Ph-positive ALL. Dasatinib is used to treat a certain type of chronic myeloid leukemia (CML; a type of cancer of the white blood cells), including treatment in people who can no longer benefit from other medications for leukemia including imatinib (Gleevec) or who cannot take these medications
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FORMULATION AND EVALUATION OF DASATINIB TABLETS because of side effects. Dasatinib is also used to treat a certain type of acute lymphoblastic leukemia (ALL; a type of cancer of the white blood cells) in people who can no longer benefit from other medications for leukemia or who cannot take these medications because of side effects. Dasatinib is in a class of medications called kinase inhibitors. It works by blocking the action of an abnormal protein that signals cancer cells to multiply. This helps stop the spread of cancer cells. Dasatinib illustrates the potential for targeted drug development based on an understanding of the genetic alterations leading to CML and the development of resistance to treatment. Observed mutations in the amino acid sequence of BCR-ABL cause the failure of treatment with existing TK inhibitors. Dasatinib has shown in vitro and in vivo activity against BCRABL, including mutations that are resistant to other available TK inhibitors. Preliminary results are available from several noncomparative studies of dasatinib in patients who were unable to tolerate or were resistant to previous therapies. The 5 phases of START (SRC/ABL Tyrosine kinase inhibition Activity Research Trials of dasatinib) represent the largest and most comprehensive evaluation of dasatinib in the treatment of patients in all stages of CML or Philadelphia chromosome-positive ALL who had undergone previous treatment for leukemia. Dasatanib had the greatest benefit in patients in the chronic phase of CML, with complete hematologic responses in 90% of patients, 52% of whom achieved a major hematologic response. Compared with those in the chronic phase, patients in the accelerated phase or blast crisis of CML, or with Philadelphia chromosome-positive ALL had lower responses. In the START-R trial, which compared the response to dasatinib and high-dose imatinib (800 mg/d), both regimens had comparable ability to induce a complete hematologic response (95% and 93%, respectively), although more patients achieved a major cytogenetic response with dasatinib (32% vs 7%). Adverse effects include significant myelosuppression. Dasatinib may have the potential for use in the management of nonleukemic malignancies. The drug is named after one of the inventor chemists, Jagabandhu Das, who was a member of the large discovery and development team at Bristol Myers Squibb.[1]

DRUG PROFILE Dasatinib is a kinase inhibitor.

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FORMULATION AND EVALUATION OF DASATINIB TABLETS The chemical name for dasatinib is N-(2-chloro-6-methylphenyl)-2-[[6-[4-(2hydroxyethyl)-1-piperazinyl]-2-methyl-4-pyrimidinyl]amino]-5-thiazolecarboxamide, monohydrate. The molecular formula is C22H26ClN7O2S H2O, which corresponds to a formula weight of 506.02 (monohydrate). The anhydrous free base has a molecular weight of 488.01. Dasatinib has the following chemical structure:

Dasatinib is a white to off-white powder. The drug substance is insoluble in water and slightly soluble in ethanol and methanol

Pharmacokinetics
Absorption
Maximum plasma concentrations (Cmax) of dasatinib are observed between 0.5 and 6 hours (Tmax) following oral administration.

Dasatinib exhibits dose proportional increases in AUC and linear elimination

characteristics over the dose range of 15 mg to 240 mg/day. The overall mean terminal half-life of dasatinib is 35 hours. Data from a study of 54 healthy subjects administered a single, 100-mg dose of dasatinib 30 minutes following consumption of a high-fat meal resulted in a 14% increase in the mean AUC of dasatinib. The observed food effects were not clinically relevant.

Distribution
In patients, dasatinib has an apparent volume of distribution of 2505 L, suggesting that the drug is extensively distributed in the extravascular space. Binding of dasatinib and its active metabolite to human plasma proteins in vitro was approximately 96% and 93%, respectively, with no concentration dependence over the range of 100500 ng/mL.
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FORMULATION AND EVALUATION OF DASATINIB TABLETS

Metabolism
Dasatinib is extensively metabolized in humans, primarily by the cytochrome P450 enzyme 3A4. CYP3A4 was the primary enzyme responsible for the formation of the active metabolite. Flavin-containing monooxygenase 3 (FMO-3) and uridine diphosphate-

glucuronosyltransferase (UGT) enzymes are also involved in the formation of dasatinib metabolites. The exposure of the active metabolite, which is equipotent to dasatinib, represents approximately 5% of the dasatinib AUC. This indicates that the active metabolite of dasatinib is unlikely to play a major role in the observed pharmacology of the drug. Dasatinib also had several other inactive oxidative metabolites. Dasatinib is a weak time-dependent inhibitor of CYP3A4. At clinically relevant concentrations, dasatinib does not inhibit CYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, or 2E1. Dasatinib is not an inducer of human CYP enzymes.

Elimination
Elimination is primarily via the feces. Following a single oral dose of [14C]-labeled dasatinib, approximately 4% and 85% of the administered radioactivity was recovered in the urine and feces, respectively, within 10 days. Unchanged dasatinib accounted for 0.1% and 19% of the administered dose in urine and feces, respectively, with the remainder of the dose being metabolites.

Mechanism of Action
Dasatinib, at nanomolar concentrations, inhibits the following kinases: BCR-ABL, SRC family (SRC, LCK, YES, FYN), c-KIT, EPHA2, and PDGFR. Based on modeling studies, dasatinib is predicted to bind to multiple conformations of the ABL kinase. In vitro, dasatinib was active in leukemic cell lines representing variants of imatinib mesylate sensitive and resistant disease. Dasatinib inhibited the growth of chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL) cell lines overexpressing BCR-ABL. Under the conditions of the assays, dasatinib was able to overcome imatinib resistance resulting from BCR-ABL kinase domain mutations, activation of alternate signaling pathways
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FORMULATION AND EVALUATION OF DASATINIB TABLETS involving the SRC family kinases (LYN, HCK), and multi-drug resistance gene overexpression.

SIDE EFFECTS INCLUDE

muscle pain weakness joint pain pain, burning or tingling in the hands or the feet skin rash skin redness peeling skin swelling, redness and pain inside the mouth mouth sores diarrhea nausea vomiting constipation stomach pain or swelling loss of appetite weight loss

Some side effects can be serious these include

fever, sore throat, chills, and other signs of infection swelling of the eyes, hands, arms, feet, ankles or lower legs sudden weight gain difficulty breathing, especially when lying down coughing up pink or bloody mucus dry cough chest pain that gets worse when coughing, sneezing or breathing deeply chest pressure dizziness fainting rapid, irregular, or pounding heartbeat
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FORMULATION AND EVALUATION OF DASATINIB TABLETS

headache tiredness confusion unusual bruising or bleeding black and tarry stools red blood in stools bloody vomit vomiting material that looks like coffee grounds slow or difficult speech weakness or numbness of an arm or leg

Dasatinib may cause other side effects

CONCLUSIONS:
Dasatinib has a wider spectrum of activity against a broader range of BCR-ABL forms than existing TK inhibitors. It has shown clinical benefit and tolerability in patients in all phases of CML, as well as in those with Philadelphia chromosome-positive ALL. Dasatinib illustrates the potential for targeted drug development based on an understanding of the genetic alterations leading to CML and the development of resistance to treatment.

REFERENCES
1. NIOSH Alert: Preventing occupational exposures to antineoplastic and other hazardous drugs in healthcare settings. 2004. U.S. Department of Health and Human Services, Public Health Service, Centers for Disease Control and Prevention, National Institute for Occupational Safety and Health, DHHS (NIOSH) Publication No. 2004 165. 2. OSHA Technical Manual, TED 1-0.15A, Section VI: Chapter 2. Controlling Occupational Exposure to Hazardous Drugs. OSHA, 1999,

http://www.osha.gov/dts/osta/otm/otm_vi/otm_vi_2.html. 3. American Society of Health-System Pharmacists. ASHP guidelines on handling hazardous drugs. Am J Health-Syst Pharm. (2006) 63:11721193. 4. Polovich M, White JM, Kelleher LO (eds). 2005. Chemotherapy and biotherapy guidelines and recommendations for practice (2nd ed). Pittsburgh, PA: Oncology Nursing Society.
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