Liposome-encapsulated Doxorubicin Research Articles

Liposome-encapsulated doxorubicin plus paclitaxel and trastuzumab as neoadjuvant chemotherapy for operable and locally advanced breast cancer.

e11069 Background: Schedules with anthracyclines and taxanes are one of the best options for primary chemotherapy. The addition of trastuzumab showed an impressive percentage of pathological complete responses in Buzdar trial (66.7%). Recently, liposome- encapsulated doxorubicin (Myocet) has been widely used in advanced breast cancer with high response rates (92% in Cortes study). The aim of our study was to assess clinical and pathological responses with liposome-encapsulated doxorubicin plus paclitaxel (and trastuzumab in pts with Her2 overexpression). Methods: 30 patients entered the study: 8 locally advanced and 22 operable. Median age was 57 years (range: 31-73). 23 pts without Her-2 3+ expression (or FISH not amplified) were treated with liposome-encapsulated doxorubicin 50 mg/m2 every three weeks for 3 courses and weekly paclitaxel 80 mg/m2 (Taxol) for 8 courses. 7 pts with Her-2 3+ expression or FISH amplified were treated with the same schedule plus trastuzumab (Herceptin) 4 mg/kg for the first administration and 2 mg/Kg for the following 7 weekly administrations. Results: Objective clinical responses were documented in 18 out of 24 evaluable pts (75%): 7 complete (29%), 7 partial (29%) and 4 minimal responses (17%). Pathological complete response was not documented in any pts. Clinical response in pts with HER-2 pverexpression treated with trastuzumab and chemotherapy was 100% (4 complete and 3 partial responses). Conservative surgery was performed in 8 out of 14 operable pts (57%) and mastectomy in 11 pts. Main toxicity was neutropenia: febrile in 3 pts (10%) and gr. 3-4 in 10 (34%). Other grade 3 toxicities were as follows: vomiting in 2 pts, asthenia in 1 pt. Conclusions: Paclitaxel and liposome-encapsulated doxorubicin was active in terms of clinical response and conservative surgery for pts with potentially operable and locally advanced breast cancer. Our study confirmed high activity of the combination with trastuzumab in pts with Her2 overexpression. However, these results in terms of clinical responses didn't turn into pathological responses. No significant financial relationships to disclose.

35 High pathologic complete remission rate with liposome-encapsulated doxorubicin + paclitaxel + trastuzumab as primary treatment in HER-2 positive operable breast cancer: clinical experience

35 High pathologic complete remission rate with liposome-encapsulated doxorubicin + paclitaxel + trastuzumab as primary treatment in HER-2 positive operable breast cancer: clinical experience

Cyclophosphamide, Vincristine, Myocet and Prednisone ± Rituximab (COMP±R) Regimen Is Safe and Effective as First Line or Salvage Therapy in Elderly Non Hodgkin Lymphoma (NHL): Preliminary Data of Single Centre Experience.

Abstract 3747Poster Board III-683 Backgrounddespite the prognosis of aggressive non-Hodgkin Lymphoma has considerably improved over the past decades, the treatment of elderly patients with NHL is still a difficult challenge for the clinician.A retrospective EORTC study conducted in patients with aggressive NHL and age above 70 years showed that about half of patients received an aggressive treatment and that only 15% of investigators employed full-dose regimens from the first cycle.We here present the preliminary data of CHOP-like regimen delivered as first-line or salvage therapy to elderly or young patients with NHL and not eligible for more aggressive therapy. Patients and methodsfrom July 2006 to January 2009, 27 pts (M/F:11/16) with a median age of 71 years (range: 53-84) were included in the study.Twenty-four pts (88%) were more than 65 yo, 18 (66%) had high-grade and 9 (34%) an indolent lymphoma. Stage III-IV disease according to Ann Arbor staging occurred in 18 pts (66%) whereas aaIPI, evaluated only for pts with aggressive NHL, was 3 2 in 9 pts (18%). ENS involvement ≥ 1 was present in 11 (41%) and BM involvement in 13 pts (48%). Most of pts (60%) had ECOG PS ≥ 1. Twenty out of 27 pts (74%) received COMP±R as first-line treatment and 7 (26%) as salvage therapy; all but one of pre-treated pts had received more than 1 line of chemotherapy (range 2-4). Twenty-five (92%) pts had co-morbidity with more than 1 disease in 44% of cases. Median LVEF was 59% (range: 35-80%). COMP±R regimen consisted of cyclophosphamide 750 mg/m2 IV d1, vincristine 1.4 mg/m2 IV d1 (capped at 2mg), liposome-encapsulated doxorubicin (MyocetÒ) at the dose of 50 mg/m2 IV d1 and Prednisone 100 mg/die PO d 1-5 with or without Rituximab at dose of 375 mg/m2 d8 at first course and at d1 of subsequent courses according to B or T-cell lymphoma phenotype respectively. To pts with early stage disease were planned 4 courses of COMP±R±IF-RT while those with advanced stage of disease received six courses of chemotherapy delivered every 3 weeks. Median number of cycles delivered was 5.6 (range: 4-8). All patients completed the planned treatment and most of them (92%) received G-CSF at dose of 300 mg/die as primary (67%) or secondary (25%) prophylaxis. ESA support was need in 8 pts (30%). Resultscomplete response (CR) was achieved in 21 (78%) and partial response (PR) in 3 (11%) of pts with an ORR of 89%; three pts had stable (n=2) or progressive disease (n=1). The regimen was safe and well tolerated with dose reduction occurring in 9 pts (34%). None of pts developed cardiac toxicity. The mainly adverse events recorded was neutropenia occurring in 15% of pts and febrile neutropenia in 6 pts (23%). Extra-haematological toxicity was mild (WHO grade 1-2) and recorded in 18% of pts. There was no treatment-related fatal toxicity. With a median follow-up of 15 months (range 6-33) the OS and EFS was 93% and 89% respectively. ConclusionCOMP±R regimen shows to be safe and effective in a group of elderly patients both as first line or salvage therapy. However more patients and longer follow-up is required to assess definitively the role of this regimen in elderly patients with untreated aggressive NHL. A prospective phase II trial is ongoing at our Institution.Tab. 1Response According to HistologyDLBCL (n=12)MZL (n=6)MCL (n=3)PTCL (n=3)FL (n=2)B-CLL (n=1)CR (%)11 (90)4 (66)2 (66)2 (66)2 (100)0PR (%)01 (16)1 (34)1 (34)00ORR %90%821001001000Tab. 2Response According to Status at TreatmentFirst line (n=20)Relapsed (n=7)CR (%)16 (80)5 (71)PR (%)3 (15)0ORR %9571 Disclosures:No relevant conflicts of interest to declare.

Biweekly Rituximab, Cyclophosphamide, Vincristine, Non-Pegylated Liposome-Encapsulated Doxorubicin (Myocet®) and Prednisone (R-COMP-14) as Induction Treatment in Poor R-IPI Risk Elderly Patients with Diffuse Large B-Cell Lymphoma (DLBCL) and Pre-Existing Heart Disease.

Abstract 1664Poster Board I-690 BackgroundDelivering of dose-dense R-CHOP represents a critical factor for improving response rate and survival outcomes in poor risk DLBCL patients (pts) but it may turn a very challenging goal to achieve in elderly pts with severe cardiac comorbidities. In this subset of pts, tachyarrhythmias, conduction disturbances, ventricular dysfunction, and a pericarditis-myocarditis syndrome, may occur during or immediately after the administration of doxorubicin hydrochloride, independently from total cumulative dose. This may lead to early treatment discontinuation and even death, strongly hampering the chance of receiving adequate chemotherapy. We have conducted a pilot study first exploring the feasibility of a dose-dense biweekly R-CHOP-like regimen including non-pegylated liposome-encapsulated doxorubicin in elderly pts at a high risk for both disease presentation and cardiac conditions. Liposomal anthracyclines, beyond the potential of a more selective uptake by lymphoma cells, may broaden the feasibility of a dose-effective treatment through a reduction of early and late cardiotoxicity. Patients and MethodsUntreated pts with poor R-IPI risk DLBCL and moderate to high ‘life threat' impact cardiopathy (NIA/NCI index) were scheduled to receive six courses of a R-COMP-14 regimen in which an equal dose (50 mg/sqm) of non-pegylated liposome-encapsulated doxorubicin (Myocet®) was substituted for doxorubicin hydrochloride within the R-CHOP platform. Treatment was given every two weeks with G-CSF support. Responses and endpoints were evaluated according to the International Harmonization Project with treatment failure defined as discontinuation of R-COMP-14 for any reason (e.g., disease progression, toxicity, initiation of new treatment without documented progression, or death). ResultsThirty-seven pts, median age 72 years (r, 62-82) with pre-existing heart disease [history of myocardial infarction (n=10), atrial fibrillation (n=8), valve disease (n=6), conduction disturbances (n=9), hypertensive cardiomyopathy (n=4)] and R-IPI score ≥ 3, were accrued in this prospective study. Thirty-three pts were in advanced stage [stage IV (n=25) + stage III (n=8): 89%], 18 (49%) had B symptoms, 29 (78%) displayed abnormal LDH and 11 (30%) presented involvement of > 2 extra nodal sites. According to Charlson, median Comorbidity Index and Score were 5.5 (r, 2-8) and 8.5 (r, 5-11), respectively. Pre-treatment left ventricular ejection fraction (LVEF) ranged between 42% and 67% (median, 57%). One-hundred eighty-four total courses were delivered (median 6, range,1-8). The median time to recycling was 15.9 days (r, 13-29) and 67% of pts received the 6 scheduled courses. Hematologic toxicity was tolerable, with CTCAE v3.0 G3/G4 thrombocytopenia and G3/G4 infections being recorded in 6 (16%) 5 (12%) pts, respectively. Seventeen failures were documented; they were due to progression (n=2), early death (n=3), treatment discontinuation (n=6) and disease recurrence (n=6). Early deaths occurred at day +12, +13 of the 1st course and day + 18 of the 2nd course, accounting for a treatment mortality rate of 8%. Treatment was discontinued because of a > 15% reduction in LVEF occurring at the 3rd and 4th courses (n=3) and the occurrence of symptomatic arrhythmia (n=3). The overall response rate was 71 % (95% CI: 53-84) with 24 complete responses and 2 partial responses. Time to Treatment Failure and Time to Progression at 28 mo.s were 51% and 63%, respectively. Disease Free Survival was 69% at a median follow-up of 14 mo.s. DiscussionThe present study documented the attractive therapeutic potential of the R-COMP-14 regimen in the setting of poor-risk elderly DLBCL pts with moderate and high impact cardiac comorbidity, in terms of both response rate and time to progression. Survival endpoints were more than satisfactory in a such unfavourable cohort of subjects for which treatment recommendations are still controversial, also due to lack of international guidelines on cardioprotective agents and strategies. Although cardiac events have occured in a fraction of pts, very early and independently from cumulative doses, they did not substancially affect the feasibility of the regimen, provided that a close clinical monitoring was ensured. R-COMP-14 represents an active dose-dense therapeutic platform for pts with cardiac comorbidity. DisclosuresNo relevant conflicts of interest to declare.

Low-Dose Gemcitabine Efficacious in Three Patients With Tumor-Stage Mycosis Fungoides

Mycosis fungoides is the most common subtype of mature T-cell lymphoma that primarily arises in the skin. The tumor manifests as patches, plaques, tumors, or erythroderma and can secondarily involve lymph nodes, peripheral blood, and visceral organs. In advanced tumor stage, chemotherapy is a second-line approach, which is generally not considered curative. Initially, most patients profit from this treatment, but observed remissions usually do not exceed several months. Because of possible immunosuppressive effects in vulnerable patients, the overall benefit of chemotherapy itself is not unequivocal in cutaneous T-cell lymphoma. We report 3 patients whose tumor-stage mycosis fungoides was not sufficiently controlled by several preceding systemic therapies, including liposome-encapsulated doxorubicin. All patients experienced excellent improvement with 250 mg/m2 gemcitabine weekly, a low-dose regimen that is predicted to be less toxic compared with standard-dose gemcitabine.

Simultaneous analysis of liposomal doxorubicin and doxorubicin using capillary electrophoresis and laser induced fluorescence

A method based on a capillary electrophoresis with laser induced fluorescence detection was developed and validated for simultaneous separation of doxorubicin (DOX) and liposomal encapsulated DOX. The separation was accomplished using a fused silica capillary (60 cm in total length, 75 μm I.D.) and potassium phosphate buffer [12.5 mM, pH 7.4] as the running buffer. The effect of sample preparation conditions on maintaining liposomal integrity was also investigated. The limit of detection for DOX was 0.1 μg/ml and the precision and accuracy of CE/LIF method was within the ranges of FDA guidelines. The validated method was successfully used to quantify DOX in human plasma using a direct injection of a 4-fold dilution of spiked liposomal DOX in human plasma.

Phase I clinical trial of liposomal-encapsulated doxorubicin citrate and docetaxel, associated with trastuzumab, as neo-adjuvant treatment in stages II and IIIA, HER2-overexpressing breast cancer patients. GEICAM 2003-03 study

BackgroundWe carried out a phase I clinical trial to establish the dose-limiting toxicity (DLT) and the maximum tolerated dose (MTD) of the combination of liposome-encapsulated doxorubicin citrate (LD) and docetaxel in breast cancer patients. Patients and methodsPatients with HER-2-overexpressing stages II and IIIA breast cancers were treated in different dose cohorts of three patients. The MTD cohort was expanded up to six patients. The patients received LD and docetaxel every 21 days, plus weekly trastuzumab, with pegfilgrastim support. ResultsA total of 20 patients were enrolled, 18 of them being assessable for toxicity and response. DLTs observed for this combination were diarrhea, fatigue, febrile neutropenia, stomatitis, myalgia, and nonneutropenic infection (pneumonia). LD 50 mg/m2 and docetaxel 60 mg/m2 every 21 days have been the MTD, with no episode of DLT observed. Seven patients developed left ventricular ejection fraction decline (six grade 1 and one grade 2). No interruptions of the treatment were needed as a consequence of cardiac toxicity. Pathologic complete response was achieved in eight patients (44%). ConclusionsThe MTD and recommended dose for phase II trials of LD and docetaxel are 50 and 60 mg/m2, respectively. The achieved results on cardiotoxicity are promising.

The safety of dose-dense liposomal-encapsulated doxorubicin in association with docetaxel (MyTax) in breast cancer.

Abstract Abstract #2157 Background: Liposomal-Encapsulated Doxorubicin (LED) shows equivalent efficacy, better cardiac tolerability at higher cumulative dose than conventional anthracyclines in breast cancer treatment.
 Methods: Sixteen pts were enrolled in a dose-finding study of LED (TLC-D99 Myocet ®) associated to Docetaxel (TXT). Twelve pts were treated with a fixed TXT dose (50 mg/m2) and TLC-D99 at three dose levels, 40-45-50 mg/m2, days 1 and 15 every 2 weeks using an intra- and inter-patient approach; four pts wrere treated at the TLC-D99 recommended dose (50 mg/m2). Cardiac monitoring of LVEF was performed every two cycles; Precursor Brain Natriuretic Peptide (proBNP) and cardiac Troponin (c-TnI) before and after 24 h chemotherapy was evaluated.
 Results: Breast cancer (BC) disease extension: metastatic (MBC), 8; locally advanced BC, 5; T2-T3 BC, 3. Previous chemotherapy: untreated, 11 pts; adjuvant, 5 pts. Enrolled pts for each dose-level: I, 7; II, 9; III, 14. Newly treated pts: I dose-level, 7; II dose-level, 3; III dose-level, 6. Valuable cycles for each dose-level in a total 77 cycles: I, 14; II, 21; III, 42. DLTs were observed in 3 pts, 21%, and 3 cycles, 4%: 2 cardiac, characterized by a 19% LVEF decrease and a symptomatic arrhythmia; one G4 hematologic resistant to G-CSF. DLTs for each dose-level by pts and cycles, respectively: I, 14% (1/7 pts) and 7% (1/14 cycles); II, no DLT in 9 pts and 21 cycles; III, 14% (2/14 pts) and 5% (2/42 cycles). Cumulative G3-4 toxicities by pts and cycles, respectively: cardiac arrhythmia 6% and 1,3%, cardiac general (symptomatic LVEF decrease), 6% and 1,3%; alopecia 81% and 65%; neutropenia resistant to G-CSF, 6% and 1,3%. Cardiac DLTs were observed in 2 elderly pts (>65 y). The 2 cardiac DLTs were observed in 2 out of 3 pts with pre-existing diastolic dysfunction. No pathologic increase of c-TnI levels was detected. Seven pts showed increased pro-BNP after chemotherapy; 1 of these with increased pro-BNP after chemotherapy, persistent the day 1 of each subsequent chemotherapy showed a DLT;
 G2 toxicities by patients and cycles, respectively: asthenia 37% and 18%, stomatitis/mucositis 12% and 5%, nausea 31% and 12%. Median rDI of TLC-D99 was 25 mg/m2/w and TXT 25 mg/m2/w for pts, respectively. Preliminary efficacy in 16 assessable pts: LA-BC and MBC, 1 CR (pCR) 7 PR (OR 62%), 4 SD and 1 PD; T2-T3 BC, 2 PR and 1 SD.
 Conclusion: dose-dense TLC-D99/Docetaxel association can be safely recommended at the dose of 50 mg/m2 for each drug. Docetaxel intensification is ongoing. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2157.

Doxorubicin pharmacokinetics following two different liposomal formulations (CAELYX™ vs MYOCET™) in advanced breast cancer patients.

Abstract Abstract #2128 Background: Liposomal encapsulation of Doxorubicin (DOX) was designed to minimize healthy tissue distribution by altering pharmacokinetics (PK) thus reducing cardiotoxicity while preserving antitumour efficacy. Study objective was to compare the PK behaviour of the two main liposomal formulations of DOX, i.e. pegylated (Caelyx, C) versus non pegylated (Myocet, M), in a single center study including 17 MBC patients, all Anthracyclin pretreated with additional risk faktors.
 Methods: Cohort 1 consisted of 10pts (mean age 62, 49-77) treated by C as 1HR infusion of 50mg/m²/4 wks. Plasma samples were collected at timepoints min 0, 30, 60, 120, 180 and d1, 7, 14, 28. In cohort 2, 7 pts (mean age 58, 51 – 70) received M 75mg/m² 1H Infusion / 3wks, plasma levels determined at min 0, 30, 60, 90, 120, 180 and d1, 2, 3. Total amount of DOX was determined following the 1st infusion respectively, analytic procedure included solid phase extraction quantification by HPLC and PK analysis by Win Nonlin Pro. Doxorubicinol was detected after M only.
 Results: in Cohort1 after C, mean Cmax of 6.8 µg/ml surprisingly occured at a mean tmax of 2.3 d, log conc time curve showed linear decrease of DOX conc suggesting continuous release into the tissue (mean conc d 7 3.3, d 14 1.0, d 21 0.3, d 28 0.09 µg/ml) . No metabolites were found, AUC (does not represent reliable drug exposure and bioavailability of DOX) expressed as µg/ml.d was 54.6. In contrast AUClast of M was calculated as 35µg ml.H and mean Cmax by 7.5 µg/ml at infusion end. DOXol was measured in all M treated patients at a slower metabolisation rate and late appearance compared to conventional DOX. t/2 el of C was 6 times longer than that of M (HR 90 vs 15). Differences in plasma disposition and PK of M vs C are depicted in table 1 and compared to pooled data of conventional DOX derived from former studies (50 mg/m² 30min inf, n=40). Cltot of conv. DOX was 10 fold higher than M and 500 fold higher than C, Vss about 12 times that of M and 70 times that of C. These PK differences underlines a different toxicity profile between PEGL (C, PPE syndrome) and NPEGL (M, myelotox, nausea), both reducing cardiac tox by avoiding peak plasma levels due to prolonged circulation time.
 
 Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2128.

Liposome-Encapsulated Doxorubicin Is Highly Active in Patients with B- as Well as T/NK-Cell Lymphomas and Cardiac Comorbidity or Older Age

Conventional anthracyclines may have severe dose-limiting side effects in patients with pre-existing cardiac comorbidity, particularly if they are elderly. In routine practice, this will lead to dose reductions or preclude the use of anthracyclines. This may result in lower remission and shorter lymphoma-free or overall survival rates. Non-pegylated liposome-encapsulated doxorubicin (Myocet®) in combination with Rituximab, cyclophosphamide, vincristine and prednisone (R-COMP) has been shown to be effective with a low cardiotoxicity profile in patients with diffuse large B-cell lymphoma (DLBCL) (Rigacci et al., Hematol Oncol. 2007; 25:198–203). We have treated 37 lymphoma patients with pre-existing cardiac disorders or elderly patients with reduced general condition who were considered ineligible or high risk for anthracycline containing therapy. Thirty seven patients with various histologies were included: 20 DLBCL, 9 T-/NK cell lymphomas, 3 follicular lymphomas, 2 post-transplant lymphoproliferative disease (PTLD), 2 chronic lymphocytic leukemia (CLL) and 1 multiple myeloma (MM). Reasons for the use of liposome-encapsulated doxorubicin were: pre-existing cardiac disorders (N=19) including myocardial infarction, cardiomyopathy, reduced left ventricular ejection fraction LVEF (N=6), elevated B-type natriuretic peptide (BNP) levels (N=10, median 680 pg/ml), aortic valve problems, or heart transplantation; older age (N=12; median age 78 years); previous therapy with anthracyclines (N=3); other comorbity or reduced general condition (N=4). Twenty-nine patients (78%) were previously untreated. The following combination regimens were given: R-COMP (N=25), COMP (N=11), and BMD (N=1). The median number of Myocet® containing cycles administered was 5. For various reasons, liposome encapsulated doxorubicin was reduced to a dose of less than 50mg/m2 in 19 patients. High remission rates were observed for this poor risk population: Complete remission rates were 70% (14/20) for DLBCL and 57% (4/7 evaluable patients) for T-/NK cell lymphomas. 31 patients (83%) are still alive at 2–9 months after the end of therapy. Five patients died from progressive disease, one of unknown reasons. Thirteen patients with DLBCL are in continuous CR 3 months after the end of therapy (range 1–8 months). No major cardiac or gastrointestinal toxicity was observed. Of note, paravasation of liposome-encapsulated doxorubicin occurred in 2 patients, but resulted in mild inflammation only without tissue damage. Hematologic toxicity was comparable to that of conventional anthracycline containing regimens: CTC Grade 3 or 4 toxicity for leukocytopenia/neutropenia occurred in 22 patients (59%). Nineteen patients (51%) received G-CSF for primary or secondary prophylaxis in at least one cycle of treatment. We conclude that liposome-encapsulated doxorubicine is highly active in combination chemotherapy for B-cell lymphomas with low cardiac toxicity in patients with pre-existing cardiac disorders or older age. Moreover, we herewith report for the first time efficacy in T/NK-cell lymphomas in this high-risk population. A randomized study evaluating cardiac effects of substitution of conventional doxorubicin by liposome-encapsulated doxorubicin in combination chemotherapy (R-CHOP vs. R-COMP) is currently being conducted by the Austrian Cooperative Group on Cancer Drug Therapy (AGMT).

Liposomal-Encapsulated Doxorubicin (Myocet™; D-99) and Vinorelbine in Previously Treated Metastatic Breast Cancer Patients: A Feasibility Study

We conducted a feasibility study to determine the safety and efficacy of liposome-encapsulated doxorubicin (Myocet) and vinorelbine in previously treated metastatic breast cancer patients. Liposome-encapsulated doxorubicin (30 mg/m2) plus vinorelbine (25 mg/m2) on days 1 and 8, every 3 weeks were given until disease progression, sever toxicity or up to 9 cycles. All patients underwent tumor assessment before enrollment. Patients with a life expectancy longer than 3 months and measurable or assessable disease were eligible. Twenty-one patients were included. Median number of treatment cycles was 5 (range, 3-9). No complete response was obtained. Stable disease and/or partial response was obtained in 9 patients. Fifteen patients experienced grade 3-4 leukopenia. There was no significant decline in cardiac function. Non-hematological toxicity was tolerable (grade 1-2). The association of doxorubicin and vinorelbine has been shown to be feasible in previously treated advanced breast cancer patients. Its efficacy should be tested as first-line therapy in metastatic patients with cardiac co-morbidities.

Improved HPLC method for doxorubicin quantification in rat plasma to study the pharmacokinetics of micelle‐encapsulated and liposome‐encapsulated doxorubicin formulations

An improved simple, rapid and accurate HPLC method for quantification of doxorubicin derived from micelle-encapsulated or liposome-encapsulated doxorubicin formulation in rat plasma was described. The mobile phase consisting of a mixture of methanol-water [containing 0.1% formic acid anhydrous and 0.1% ammonia solution (25%), pH 3.0], 60:40, was delivered at a flow rate of 1.0 mL/min. Sample preparation for micelle- or liposome-encapsulated doxorubicin in rat plasma were achieved directly by protein precipitation with acetonitrile. Doxorubicin and daunorubicin (internal standard, IS) were separated on a C(18) reversed-phase HPLC column and quantified by a fluoresence detection with an excitation wavelength of 475 nm and an emission wavelength of 580 nm. The linearity was obtained over the range of 5.0-1000.0 ng/mL and 1.0-200.0 microg/mL for doxorubicin and the lower limit of quantitation was 5.0 ng/mL. For each level of quality control samples, inter- and intra-assay precision was less than 9.6 and 5.1% (relative standard deviation), respectively, and percentage error was within +/-2.6%. The extraction recoveries of doxorubicin in the range of 10 ng/mL to 100 microg/mL in rat plasma were between 94.1 and 105.6%. This method was successfully applied to the pharmacokinetic study of doxorubicin formulations after i.v. administration to rats.

Pharmacokinetics of intra-arterial applied liposomal encapsulated doxorubicin in liver metastases or hepatocellular carcinoma: A phase I study

13523 Background: Doxorubicin has been one of the most widely used anti-neoplastic agents as a single-agent systemic therapy for HCC. Doxorubicin is also used in locoregional therapy approaches. Caelyx (Doxil) is a doxorubicin formulation of polyethylene glycol-coated liposomes. For this new form of doxorubicin, a superior pharmacokinetic profile over that of free doxorubicin is postulated. Blood flow reduction by micro embolization with degradable starch microspheres (DSM; e.g. Spherex, EmboCept) is effective in HCC and liver metastasis. Purpose: Evaluation of the pharmacokinetics (PK) of the encapsulated as well as the released fraction of doxorubicin in addition to the determination of the maximum tolerated dose of intra-arterially applied liposomal encapsulated doxorubicin (Caelyx) in HCC and liver metastases in combination with DSM. Methods: All patients with histologically proven HCC or liver metastases had shown progressive disease when previously treated with systemic chemotherapy. Six dose escalation steps from 15 mg/m2 to 100 mg/m2 were performed with two or three patients in each dose group. Each patient received one single application of Caelyx/DSM. The distribution of Caelyx depended on the tumor size in the left and right liver lobe. For the quantification of Caelyx, doxorubicin, and doxorubicinol, a validated RP-HPLC method with fluorometric detection (excitation wavelength: 470 nm, emission wavelength: 550 nm) was used. Results: PK was calculated for 13 out of 14 treated patients. The mean (terminal) half life for Caelyx was calculated as 3 days, mean residence time about 5 days, the Vss 1.5 l, clearance was determined to be less than 1 ml/min. The increase in peak plasma concentration as well as AUC was linear between 15 and 100 mg/m2 Caelyx. The MTD was reached at a single dose of 100 mg/m2 due to WHO- grade 3 myelosuppression in two patients without further SAE. Conclusions: These first results indicate that hepatic intra arterial chemotherapy with Caelyx in combination with DSM is well tolerated and achieved encouraging responses in patients with liver metastases or HCC. The main PK parameter for Caelyx in combination with DSM did not significantly differ from those obtained after systemic intravenous infusion. Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Expert Testimony Other Remuneration Essex Pharma GmbH, PharmaCept GmbH

Distribution of Free and Liposomal Doxorubicin After Isolated Lung Perfusion in a Sarcoma Model

Isolated lung perfusion (ILP) with free and a novel liposomal-encapsulated doxorubicin (Liporubicin, CT Sciences SA, Lausanne, Switzerland) was compared with respect to drug uptake and distribution in rat lungs bearing a sarcomatous tumor. A single sarcomatous tumor was generated in the left lung of 39 Fischer rats, followed 10 days later by left-sided ILP (n = 36) with free and equimolar-dosed liposomal doxorubicin at doses of 100 microg (n = 9) and 400 microg (n = 9) for each doxorubicin formulation. In each perfused lung, the drug concentration and distribution were assessed in the tumor and in three areas of normal lung parenchyma by high-performance liquid chromatography (n = 6) and fluorescence microscopy (n = 3). Histologic assessment and immunostaining with von Willebrand factor was performed in 3 animals with untreated tumors. The sarcomatous tumors in controls were well vascularized with fine branching capillaries present throughout the tumors. Isolated lung perfusion resulted in a heterogeneous drug distribution within the perfused lung and a consistently lower drug uptake in tumors than in lung parenchyma for both doxorubicin formulations and both drug doses applied. Isolated lung perfusion with free doxorubicin resulted in a significantly higher drug uptake than Liporubicin in both the tumor and lung tissue for both drug doses applied (p < 0.01). However, the tumor/normal tissue drug ratio was lower for free than for liposomal doxorubicin at a drug dose of 100 microg (0.27 +/- 0.1 vs 0.53 +/- 0.5; p = 0.225) and similar for both doxorubicin formulations at a drug dose of 400 microg (0.67 +/- 0.2 vs 0.54 +/- 0.2; p = 0.335). Both doxorubicin formulations resulted in fluorescence signaling emerging from all tissue compartments of normal lung parenchyma but only in weak and sporadic signaling from the tumors confined to the tumor periphery and vessels situated within the tumor for both drug doses assessed. Isolated lung perfusion with free and liposomal doxorubicin resulted in a heterogeneous drug distribution within the perfused lung and in a lower drug uptake in tumors than in lung tissue for both doxorubicin formulations and drug doses applied.

Clinical and Pharmacokinetic Characteristics of Intracavitary Administration of Pegylated Liposomal Encapsulated Doxorubicin in Dogs with Splenic Hemangiosarcoma

Background: Canine splenic hemangiosarcoma (HSA) is a fatal malignancy, and most affected dogs die within a few months of diagnosis. Most dogs present with signs from tumor rupture, resulting in hemoabdomen and intra‐abdominal dissemination. The abdomen is also the main site of disease recurrence. Hypothesis: Intraperitoneal (IP) administration of doxorubicin will delay or prevent intra‐abdominal tumor recurrence and prolong survival in dogs with HSA. Animals: Fourteen dogs with splenic HSA. Methods: A prospective, unmasked, uncontrolled clinical trial. After staging of disease status and splenectomy, pegylated liposomal encapsulated doxorubicin was administered intraperitoneally (1 mg/kg body weight) every 3 weeks for 4 cycles. All dogs were monitored for recurrence of HSA. Samples of plasma and abdominal fluid were collected for measurement of doxorubicin concentration and pharmacokinetic analysis. Nonlinear mixed‐effect modeling was used to describe the pharmacokinetics of liposomal doxorubicin administered IP. Results: All 14 dogs died, 12 because of HSA and 2 from other causes. Postmortem examination was performed on 12 dogs. All 12 dogs died because of HSA‐related causes and had hepatic metastases and hemoabdomen. The IP‐treated dogs had fewer serosal, mesenteric, and omental metastases than historical controls treated with systemic doxorubicin. Results of the postmortem examination and pharmacokinetic analysis confirmed that IP delivery of doxorubicin resulted in an effective drug concentration with a clearance comparable with that after IV delivery. Conclusions and Clinical Importance: IP pegylated liposomal encapsulated doxorubicin administration did not prevent intra‐abdominal recurrence of HSA in dogs.

Liposomal Doxorubicin in Breast Cancer

Drug-delivery carriers represent an important step in the development of targeted therapy. Encapsulation of drug into liposomes represents such a carrier, and helps to minimize side effects of conventional doxorubicin by improving the tumor-specific biodistribution profile. We review the development of the two liposomal doxorubicin formulations, pegylated liposomal doxorubicin and liposomal-encapsulated doxorubicin citrate from reconstitution and comparative pharmacokinetics to pivotal Phase III trials, with special emphasis in breast cancer. The relative differences in the toxicity profile can be attributed to their differences in the liposomal formulations. Areas of special interest include the reduction in cardiac toxicities and the improved efficacy, such as in the treatment of ovarian cancer. These improvements have also increased the potential of these liposomal formulations of doxorubicin for combination and sequencing with other biological and cytotoxic agents for clinical benefit.

Liposome‐encapsulated doxorubicin in combination with cyclophosphamide, vincristine, prednisone and rituximab in patients with lymphoma and concurrent cardiac diseases or pre‐treated with anthracyclines

To assess the efficacy and safety of the combination of non-pegylated liposome-encapsulated doxorubicin (Myocet(R)) with cyclophosphamide, vincristine, prednisone and rituximab (R-COMP) in patients with aggressive non-Hodgkin's B-cell lymphomas. Twenty-one patients were selected for the presence of negative concurrent clinical features such as cardiac comorbidity and/or previous treatment with anthracycline-based regimens. Liposome-encapsulated doxorubicin at a dose of 50 mg/m(2) was administered in association with cyclophosphamide (750 mg/m(2)), vincristine (1.4 mg/m(2)), prednisone (40 mg/m(2)) and rituximab (375 mg/m(2)) every 21 days for four to six cycles unless progression or unacceptable toxicity occurred. A complete response (CR) was obtained in 16/21 patients (76%), three patients achieved a partial response (14%), with an overall response rate of 90%. Two patients (10%) did not respond to therapy. After a median follow-up of 13 months (range 2-36 months), 2/16 CR patients relapsed, with a disease-free survival (DFS) of 78%. The replacement of doxorubicin with its non-pegylated liposomal pharmaceutical analogue was well tolerated and highly effective in inducing remission in this group of patients at high risk for cardiac toxicity or previously treated with anthracyclines. Its high tolerability and low incidence of cardiac events (only one patient) warrants further studies to confirm the clinical benefits of liposomal doxorubicin in this subset of patients.

Liposomal-encapsulated doxorubicin plus cyclophosphamide as first-line therapy in metastatic breast cancer: a phase II multicentric study

BackgroundThe objective of this study is to evaluate the efficacy and toxicity of the liposome-encapsulated doxorubicin (TLC D-99) plus cyclophosphamide (CTX) as first-line treatment of metastatic breast cancer in light of the potential cardioprotective effect of TLC D-99 as compared with conventional doxorubicin. Materials and methodsSixty-seven patients as defined according Simon's two-stage phase II design were enrolled. They received TLC D-99 at the dosage of 60 mg/m2 plus CTX 600 mg/m2, with cycles repeated every 3 weeks. Cardiac function was assessed by ultrasonography at baseline and every two cycles. ResultsThe principal characteristics of the 67 enrolled patients were as follows: median age 60 years (range 33–75), median World Health Organization performance status of 1 (range 0–2) and dominant disease site (viscera/bone/soft tissue): 47/15/15 There were nine complete responses and 32 partial responses for an overall response rate of 64%; a further 14 patients had stable disease and the remaining nine patients progressed. Median number of administered cycles was six. Median duration of response was 10 and 9 months, respectively, for complete responders and partial responders. Median duration of survival was 17+ months (range 3 to 33+). Hematological toxicity consisted in leucopenia (G1–G2) in 21 patients and anemia (G1–G2) in 20 patients; G1 thrombocytopenia was observed only in 2 patients. Non-hematological toxicity was generally mild with G1–G2 nausea/vomiting in 23 patients and G1–G2 mucositis in 10. Hair loss was registered in 30 patients and it was G2 in 14 patients. As to concern cardiac toxicity, one patient developed an asymptomatic 20% decline of left ventricular ejection fraction from the baseline value. ConclusionsThe results of our study show that the combination of TLC D-99 plus CTX is active and well tolerated, with no unexpected toxicity.

Clinical and Pharmacokinetic Characteristics of Intracavitary Administration of Pegylated Liposomal Encapsulated Doxorubicin in Dogs with Splenic Hemangiosarcoma

Canine splenic hemangiosarcoma (HSA) is a fatal malignancy, and most affected dogs die within a few months of diagnosis. Most dogs present with signs from tumor rupture, resulting in hemoabdomen and intra-abdominal dissemination. The abdomen is also the main site of disease recurrence. Intraperitoneal (IP) administration of doxorubicin will delay or prevent intra-abdominal tumor recurrence and prolong survival in dogs with HSA. Fourteen dogs with splenic HSA. A prospective, unmasked, uncontrolled clinical trial. After staging of disease status and splenectomy, pegylated liposomal encapsulated doxorubicin was administered intraperitoneally (1 mg/kg body weight) every 3 weeks for 4 cycles. All dogs were monitored for recurrence of HSA. Samples of plasma and abdominal fluid were collected for measurement of doxorubicin concentration and pharmacokinetic analysis. Nonlinear mixed-effect modeling was used to describe the pharmacokinetics of liposomal doxorubicin administered IP. All 14 dogs died, 12 because of HSA and 2 from other causes. Postmortem examination was performed on 12 dogs. All 12 dogs died because of HSA-related causes and had hepatic metastases and hemoabdomen. The IP-treated dogs had fewer serosal, mesenteric, and omental metastases than historical controls treated with systemic doxorubicin. Results of the postmortem examination and pharmacokinetic analysis confirmed that IP delivery of doxorubicin resulted in an effective drug concentration with a clearance comparable with that after i.v. delivery. IP pegylated liposomal encapsulated doxorubicin administration did not prevent intraabdominal recurrence of HSA in dogs.

Liposome-encapsulated doxorubicin citrate in previously treated recurrent/metastatic gynecological malignancies

The aim of this study was to evaluate the safety and efficacy of liposome-encapsulated doxorubicin citrate (LEDC) in patients affected by recurrent/metastatic gynecological malignancies scheduled for palliative chemotherapy. Inclusion criteria were proven recurrent/advanced gynecological neoplasms, measurable/assessable disease, adequate organ function, left ventricular ejection fraction >50% as determined by echocardiography, informed consent. LEDC was administered intravenously over 1 h at the dose of either 75 mg/m(2) or 60 mg/m(2) (every 3 weeks until disease progression or toxicity prohibiting further therapy). From May 2003 to September 2005, 36 patients were enrolled. Primary disease was ovarian, endometrial, and cervical cancers in 15 (42%), 11 (30%), and 10 (28%) patients, respectively. LEDC was employed as third- or fourth-line chemotherapy in 25 (70%) and 11 (30%) patients, respectively. The median number of courses of LEDC received was 3 (range 2-9). Six patients (17%) achieved a partial response to treatment lasting 27 weeks and 10 patients (28%) experienced stable disease lasting 18 weeks. The predominant toxicity was hematological, especially neutropenia. Among patients receiving a dose of 75 mg/m(2), two (11%) suspended therapy for febrile neutropenia, and nine (50%) required a dose reduction of 25%. As a result, the next 18 patients were treated at a reduced dose (60 mg/m(2)) of LEDC. Severe neutropenia (G3-G4) was significantly less common in this group (61% versus 22%; P= 0.04). LEDC has shown antineoplastic activity in previously treated recurrent/metastatic gynecological cancer patients and the toxicity profile could be considered acceptable at a 60 mg/m(2) dosage.