Liposomal Adriamycin Research Articles

Ixazomib, Lenalidomide, and Dexamethasone (IRd) Versus Ixazomib, Pegylated Liposomal Adriamycin, and Dexamethasone(IDd) in Elderly Frail Newly Diagnosed Multiple Myeloma with GA Score ≥2: A Prospective Multicenter Study

Introduction Frailty patients with newly diagnosed multiple myeloma (NDMM) have inferior survival and higher rate of treatment discontinuation due to complications, poor tolerance and compliance and lack of effective treatment. The aim of this study was to prospectively investigate the treatment tolerability, clinical outcomes and quality of life of the ixazomib based regimens in frail NDMM patients according to IMWG frailty index. This trail is registered at http://www.chictr.org.cn/index.aspx as chiCTR1900024917. Methods This prospective trail was an investigator initiate, multicenter, non-randomized controlled study which frail NDMM patients received 6-8 four-week induction cycles of ixazomib, lenalidomide and dexamethasone (IRd) and ixazomib, pegylated liposome adriamycin and dexamethasone (IDd), followed by matienance therapy of 4-week with ixazomib and dexamethasone (Id) until progression. Inclusion criteria were aged ≥ 65 years, IMWG GA score ≥ 2 or Mayo geriatric vulnerability scoring system defined as vulnerable. Main exclusion criteria were acute cardiovascular and cerebrovascular events. The primary endpoint was overall response rate (ORR) defined by having at least a partial response (≥PR). The secondary endpoints were progression free survival (PFS), overall survival (OS), adverse events (AE) and health-related quality of life (using the EORTC-QLQ C30). Results From July 2019 to December 2021, a total of 132 patients from 15 hospitals were enrolled of which 12 were excluded because of ineligibility and non-randomized assigned to receive IRd group (n=60) or IDd (n=60) according to doctor's estimated (Figure 1). The median age was 71 years range (65-88). All patients had a GA Index higher than 2 score, in which 27.5% were also defined fragile by Mayo criterion. The ORR was 83%, including 22 (19.6%) patients with a complete response (CR) and 45 (40.2%) with a very good partial response (VGPR). The ORR was 84.2% and 81.8%, CR was 26.3% and 17.2, VGPR was 36.8% and 43.6% of IRd and IAd group, respectively. After a median follow-up time of 22.9m (95% CI 18.9-26.9), the median PFS was 19.5m of all patients, 24.6m and 15.8 of IRd and IDd group, respectively. The median OS of all patients and IRd group was no reach, while 29.2m in IDd group (Figure 1). Four patients died in 60 days post enroll up because of pneumonia caused by bedridden. Twenty-six and thirty-six patients did not process to maintenance therapy in IRd and IAd group, due to PD (14 in IRd and 16 in IDd), AE (6 in IRd and 7 in IDd), incompliance (5 in IRd and 9 in IDd) and early death (1 in IRd and 3 in IDd). Cumulative grade 3 or higher hematological AEs occurred in 10/60 (16.7%) and 13/60 (21.7%), whereas grade ≥3 non-hematological AEs were reported in 25/60 (41.7%) and 46/60 (76.6%) patients. Most common non-hematological grade ≥3 AEs were infections (20.8%) and gastro-intestinal (13.3%). Only 1 patient experiencing PNP grade 3 (0.83%). During induction, patients experienced a statistically and clinically significant improvement in GHS/QoL of both IRd and IAd group (p<0.0001). (Figure 1) Conclusion The results of ORR and safety support the use of IDd or IRd as frontline therapy for fragile elderly MM patients, which also improves QoL. IDd was especially suitable for frail MM patients with renal dysfunction and thrombosis. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Adriamycin - efficacy and possible adverse effects

Abstract Adriamycin (doxorubicin) is a chemical substance in the anthracycline class with a wide range of applications in oncology and hematology. The mechanism of action of Adriamycin is related to formation of irregular bonds between nucleobases of DNA and inhibition of key enzymes of DNA synthesis - topoisomerase I and II as well as to formation of free radicals damaging DNA. A major limitation in the drug use is associated with its adverse effects such as cardiotoxicity and hepatotoxicity. The mechanism of myocardial injury by Adriamycin is linked to an increase in oxidative stress associated with impaired mitochondrial function and structure. Cardiotoxicity of anthracyclines is classified as: acute, chronic or late (delayed). Hepatotoxicity of Adriamycin as a damage of the liver is associated with a dysfunction of this organ. Adriamycin studies have shown increased level of transaminase present in 40% of patients treated with Adriamycin. The state was transient and asymptomatic, returning to the initial level even when treatment continued. Knowledge of cancer diseases contributed to a successive creation of two improved forms of Adriamycin (doxorubicin) – nonpegylated and pegylated formulas of the drug. The mechanism of anticancer effects of liposomal Adriamycin is similar to the mechanism of conventional Adriamycin, but placement of the molecules of active substance in liposomes has significant influence on the distribution of the drug. In order to increase the distribution of the drug, a special form of liposomal Adriamycin has been created by covering the surface of the liposomes with a hydrophilic polymer - (MPEG). This process, known as pegylation, decreases the interactions between the lipid bilayer membrane and the plasma components. Pegylated form of the drug is associated with a higher incidence of acute complications.

Reacción de hipersensibilidad a carboplatino no reversible mediante protocolo de desensibilización

Platinum compounds are widely used in different tumors, being carboplatin indicated in ovarian carcinoma. The main carboplatin-dose limiting toxicity is the thrombocytopenia, however, sometimes we have seen hypersensitivity reactions that, although it is not a fact too often (8-16%), can be severe. For this reason, desensitization protocols have been developed.We report the case of a patient intervened surgically from a papillary serous carcinoma of ovary (stage IIIC) and who later received adjuvant chemotherapy with carboplatin and paclitaxel. After twenty-two months of disease-free interval, she showed unresectable abdominal relapse, so it was again treated with carboplatin and paclitaxel. During the administration of the second cycle, said severe hypersensitivity reaction to carboplatin.Attempted to desensitization unsuccessfully, can not continue with this treatment scheme and forcing to start second line with trabectedin and pegylated liposomal adriamycin.

Local administration of liposomal adriamycin inhibited proliferation of metastatic cells in axillary lymph nodes in rabbit breast cancer model

Objective To assess the inhibitory effects of liposomal adriamycin (LADR) locally injected into mammary glands of VX2 tumor-bearing rabbits on proliferation of lymph nodal metastatic cells. Methods Twenty-one VX2 tumor-bearing rabbits were randomly and equally divided into 3 groups. Rabbits were randomized to receive sham treatment (Group I), subcutaneous LADR around tumor (Group II) and intravenous free adriamycin (Group III), respectively. Breast tumor and axillary lymph nodes were harvested after 3 repeated treatment. Nodal sizes of both pre- and post-treatment were measured. Proliferating cell nuclear antigen (PCNA) mRNA in both tumor and lymph nodes were determined by RT-PCR. Results The mean size of axillary lymph nodes in Group I, II and III increased by 3.70%, 1.55% and 2.89%, respectively, with significant difference between Group III and I ( P=0.004) and between Group II and III ( P=0.002). Relative expression values of PCNA mRNA in breast tumors of Group I, II and III were 0.486, 0.513 and 0.396, respectively. For Group III, PCNA mRNA was significantly less expressed than that in Group I ( P=0.023) and II ( P=0.005). Relative expression values of PCNA mRNA in axillary lymph nodes of Group I, II and III were 0.541, 0.329 and 0.450, respectively. Compared with Group I, Group III showed a markedly decreased expression of PCNA ( P=0.021). The least level of PCNA mRNA was found in Group II, with a significant difference from that in Group III ( P=0.004). Conclusion Local injection of LADR was an effective therapeutic regimen for lymphatic metastases from breast cancer, regardless of its little effect on primary tumor.

VAMP/ThaCyDex: Velcade® (Bortezomib), Adriamycin, Melphalan and Prednisone Alternating with Thalidomide, Cyclophosphamide and Dexametasone as a Salvage Regimen in Relapsed Multiple Myeloma Patients

Background: Bortezomib and Thalidomide have demonstrated to be effective in relapsed and refractory Multiple Myeloma (MM) patients, including those with adverse cytogenetics (CG). Moreover, Bortezomib and thalidomide-based combinations result attractive to improve efficacy, but toxicity could be increased. Based on this background, we have tested if an alternating regimen consisting on two highly effective schedules could overcome MM drug resistance without an increase in toxicity in relapsed/refractory MM patients.Patients and Methods: Treatment schedule consisted on 6 alternating cycles of VAMP (Bortezomib 1,3mg/msq IV days 1,4,8 and 11; Melphalan, 9mg/msq po, days 1–4; Prednisone 60mg/msq po, days 1–4; and conventional or liposomal Adriamycin 40 or 30mg/msq respectively on day 1 of a 28 day cycle) alternating with ThaCyDex (Thalidomide 200mg/d po day 1–28; Cyclophosphamide 50mg/d po, days 1–28; and Dexametasone 40mg/d po, days 1–4). After 6 cycles, responding patients, received the previous schedule, every other month as consolidation therapy.Results: From June 2007 until August 2008, 20 patients have been included, with a median age of 63 years (Range 48–81); 15 patients (75%) had previously received autologous stem cell transplantation. 6 patients (30%) had previously received Bortezomib-based therapy, and one patient (5%) had previously received IMID-based therapy. Efficacy and toxicity were evaluated on an intent-to-treat basis. After a median of 6 cycles, 20 patients were evaluable for response, 9 patients (42%) achieved immunofixation negative Complete Response (CR), 3 patients (16%) nCR, 9 patients (47%) partial response, which makes an ORR of 94,7%. In addition, 1 patient (5%) remained in stable disease and one patient died during induction therapy due to septic shock. Seven patients presented high risk cytogenetic abnormalities [t(4;14) and/or delRB], and CR was obtained in 3 of them (42%) plus one additional nCR in 1 patient (14%). Moreover, allogeneic stem cell transplantation was performed in two of these high risk patients, as they were effectively rescued by this salvage regimen. Two patients progressed during the consolidation treatment. Toxicity was manageable, being haematologic events the most frequently reported. 6 patients (30%) developed ≥G3 thrombocytopenia and 6 patients (30%) neutropenia. Infection ≥G3 occurred in 3 patients (16%). Despite the combination of two drugs with potential neurologic toxicity, the use of them in alternated schedule resulted in that only 3 patients (15%) developed Peripheral Polyneuropathy, none of them >G2.Conclusion: Preliminary results show that alternating VAMP/ThaCyDex is a highly effective salvage regimen in relapsed/refractory MM patients, including high risk subgroup with adverse cytogenetic abnomalities. Haematologic toxicity was the most frequent adverse event, while the incidence of Peripheral Polyneuropathy was low, despite the use of two neurotoxic drugs. This results indicate that the alternating approach allows the exposure to a large number of active drugs without increase in the toxicity. A second analysis will be performed in December 2008 and results will be updated.

Primary Sarcoma of the Prostate and Thrombocythemia in Adult; A First Case Report in English Literature

Sarcoma is an unusual type of prostate cancer in adult. Prostate sarcoma (PS) is rare with only 183 cases reported in the English language medical literature from 1938-1998. Here we present a case of african.national who presented with locally advanced prostate sarcoma with thrombocythemia. However responded favourably to his initial trearment. He was treated with anthracycline based systemic chemotherapy; monoclonal antibody/large molecular target therapy (cyclophosphamide, liposomal adriamycin, vincristine, Cetuximab) for first three months without grade 3 or 4 toxicity. Then given synchronous chemo-target-radiation therapy in view of favourable initial radiological response and clinical benefit (effective palliation in terms of pain control and bleeding control).

Local injection of liposomal adriamycin to inhibit metastatic cell proliferation in axillary nodes in rabbits bearing breast tumors

Objective To assess the inhibitory effects of local injection of liposomal adriamycin (LADR) on the proliferation of lymph node metastases in rabbits bearing VX2 carcinoma in the mammary gland. Methods Thirty female New Zealand white rabbits were divided into 3 groups, with 10 in each. VX2 tumor mass suspensions were injected into the breast tissues of rabbits. Treatment initiated once the axillary lymph node reached 5 mm in the maximum diameter. Group 1 received a sham treatment. Group 2 received a subcutaneous injection of LADR adjacent to tumor. Group 3 received an intravenous injection of free ADR (FADR) at the same dose and concentration to group 2. The breast tumors and axillary lymph nodes were resected after the treatment was repeated 3 times. The tumor and node sizes before and after treatment were measured. PCNA mRNA expressions in breast tumors and axillary nodes were determined using RT-PCR. Results The mean growth ratios of lymph nodes after treatment were 3.70, 1.55, and 2.89. respectively, in groups 1, 2, and 3. The slowest node growth was observed in animals of group 2. with significant differences from group 1 ( P<0.001) and group 3 ( P=0.002). The relative values of PCNA mRNA expression in lymph nodes were 0.541, 0.329, and 0.450, respectively, in groups 1, 2. and 3. Group 2 exhibited a significantly reduced PCNA mRNA expression in metastatic lymph node, as compared to group 1 ( P<0.001) and group 3 ( P = 0.004). Intravenous FADR injection effectively lowered the mRNA expressions of PCNA in breast tumors, which were not apparently altered after local LADR injection. Conclusion Local injection of LADR holds a strong inhibitory effect on the proliferation of metastatic tumor cells in lymph nodes and appears to be an effective method for the treatment of lymphatic metastases of breast cancer.

Estudio Fase II: doxorrubicina liposomal pegilada y ciclofosfamida en pacientes con cáncer de ovario avanzado platinorresistente

Purpose: To evaluate the efficacy and safeness of patients with advanced ovary cancer resistant to platinum-based drugs receiving a regime of liposomal adriamycin 35 mg/m 2 and cyclophosphamide 600 mg/m 2 day 1 every 28 days, up to a maximum of 8 cycles. Material y methods: Twenty two evaluable patients were enrolled using the indicated phase II methodology. Results: A response was achieved in 5 out of 21 evaluable patients (23.8%; IC: 5.6-42), with 4 PR (19%; IC: 2.3-35.8) and 1 CR (4.8%; IC: 0-13.9). Hematological toxicity was grade 1,2 anemia in 12 (59%), and 3 (4%), and grade 1-2 neutropenia (26%). Significant non-hematological toxicity was grade 3 fatigue (4%), nausea (4%), and palmoplantar erythrodisesthesia (4%). The mean time was 3.03 months (IC: 0.96-5.11). The median of survival was 6.13 months (IC: 4.32-7.95). The mean duration of the response in patients responding to treatment was 4.73 months (IC: Conclusion: The pegilated liposomal doxorubicin and cyclophosphamide regime administered to patients with advanced ovary cancer resistant to platinum-based drugs is relatively effective and shows low toxicity.

Treatment of rabbits bearing advanced VX2 tumors in the mammary gland with nano-sized liposomal adriamycin administered by various routes

To investigate the therapeutic effect of nano-sized liposomal adriamycin (NLADR) administered by various routes on rabbits bearing advanced breast tumors. NLADR with a mean diameter of 120 nm was prepared by pH gradient-driven drug encapsulation method. VX(2) tumor mass suspension was injected into the breast tissues of 50 female New Zealand White rabbits. Six weeks after the inoculation 38 surviving tumor-bearing rabbits were randomly divided into 5 groups: group A (control group), receiving a sham treatment; group B, receiving subcutaneous injection of NLADR with a dose of 1 mg/kg into the areas adjacent to the implanted tumor; group C, receiving intravenous injection of NLADR with a dose of 1 mg/kg; group D, receiving NLADR with a dose of 1 mg/kg administered by subcutaneous injection combined with intravenous injection with half of the whole dose for both routes; and group E, receiving intravenous injection of free adriamycin (FADR). The treatment was repeated every 48 hours. The rabbits were killed 48 hours after the third treatment. The breast tumors, axillary lymph nodes, and all of the metastatic tumors anatomically detected in distant organs were collected. The sizes of tumors and axillary nodes before and after treatment were measured. Necrosis of tumor tissue was assessed pathologically. The mRNA expression of proliferating cell nuclear antigen (PCNA) was determined using RT-PCR. Apoptosis was identified and quantified as apoptosis index (AI) using TUNEL method. The average growth rate of tumor was the highest in group A (1.58) and the lowest in group C (1.33). The average growth rate of axillary lymph nodes was the highest in group A (3.70), significantly higher than in any other groups (all P = 0.00); and was the lowest in group B, significantly lower than groups A, C, and E (all P < 0.01), however without a significant difference between groups B and D (P = 0.148). The PCNA mRNA expression level of the implanted tumor in groups C was the lowest, significant lowest then those in group A and B (both P < 0.01). The sequence of PCNA mRNA in the axillary lymph nodes was group B < group D < group C < group E < group A with significant differences between group B and groups A, C, and E. The sequence of PCNA mRNA in the mediastinal lymph nodes was group B < Group E < group D < group C < group A. The PCNA mRNA expression level was the lowest in group D, significantly lower than that in group A (P = 0.011). Necrosis of implanted tumor, metastatic foci in lung and liver, and lymph nodes was obvious in groups C, D, and E. Necrosis of implanted tumor and metastatic foci in lung and liver was significantly obvious in group C than in group D (P = 0.000 and P = 0.022). Necrosis of the implanted tumor was significantly more obvious in group C than in group F (P = 0.033). Necrosis of axillary lymph nodes was significantly more obvious in group than in groups C and E (P = 0.000 and P = 0.000). The values of AI of the implanted tumor in groups C and D were 16.74% and 18.04%, both significantly higher than those in group E (P = 0.02 and P = 0.04), and those of group E were significantly higher than those in groups A and B (both P < 0.01). The AI value of the axillary lymph nodes was 21.73%, significantly than those in groups A, C, and E (all P < 0.01). The average AI values of the metastatic foci were 16.52%, 15.77%, and 14.50%, all significantly higher than that in group B (all P < 0.01), and that of group B was significantly higher than that in group A (P = 0.039). NLADR, especially intravenous administration combined with subcutaneous administration, is effective in treatment of advanced breast carcinoma with lymphatic and distant metastases.

Pegylated Liposomal Doxorubicin (Doxil®), Dexamethasone and Low Dose Thalidomide (DDt) as Therapy for Newly Diagnosed Multiple Myeloma.

Background: Thalidomide with dexamethasone is considered the current standard induction therapy for multiple myeloma. Previously, treatment consisted mainly of infusional vincristine, adrimaycin and dexamethasone (VAD). High dose thalidomide is associated with significant toxicity. We conducted a study to evaluate efficacy and toxicity of adding liposomal adriamycin (Doxil®) to dexamethasone and low dose thalidomide. Stem cell harvest yield was also assessed.Methods: Patients were treated with Doxil® 40mg/m2 IV d1, thalidomide 100 mg PO Q HS, and dexamethasone 40 mg PO d1–4 and 15–18 Q 28 days for 4 cycles. After 4 cycles, eligible patients underwent autologous stem cell transplant. Patients received prophylactic Amoxicillin 250 mg PO Q day and Acyclovir 200 mg PO BID. Erythropoietin and biphosphonates were administered. Aspirin was changed to full dose coumadin anticoagulation after one death from pulmonary embolism.Results: Between 3/03 and 5/05, 11 out of planned 25 patients were enrolled on the study; 1 patient excluded for wrong diagnosis. Median age 61.5 (51–81yrs); 7 males; 7 IgG, 2 IgA and 1λ-light chain subtypes; Stage IB-1, Stage IIA-4, Stage IIIA-5; median albumin 3.75 (range:1.6–5.1 g/dL), creatinine 0.95 (range: 0.7 – 2.7 mg/dL) and B2M 1.95 (range: 1.1–10.1 mg/L). Four patients completed 4 cycles; one is undergoing treatment; three patients taken off study-one after 2 cycles per patient's request, one secondary to grade 4 toxicity after cycle 1, and another after greater than 3 weeks treatment delay after cycle 3 secondary to pneumonia. Three patients (30%) developed grade 3–4 toxicities- two due to pneumonia (after cycles 2 and 3, respectively); and one with dehydration and renal failure after cycle 1. There were 2 deaths-one secondary to pulmonary embolism after cycle 1; another due to myocardial infarction after cycle 2. Two patients developed VTE- one after cycle 1 while on low dose coumadin; another died of pulmonary embolism after cycle 1, while on aspirin, requiring amendment to full dose coumadin anticoagulation. No further thrombotic episodes were encountered. Five patients underwent stem cell harvest with cyclophosphamide mobilization. The median harvest yield was 9.76 x106 CD34/kg (range: 5.27 – 31.94) with a median of 1 day (range: 1–7) to achieve at least 5 x 106 CD34/kg. Of the ten patients who received at least one cycle and assessable for response, the ORR was 80% (near complete remission(nCR)-10%; PR-70%), 1 MR, 1 SD. The M-spike response after first cycle of therapy ranged from 8% to 71%, with six patients (60%) having greater than 50% reduction in M-protein level. One patient achieved nCR after 2 cycles. Responses after the 3rd and 4th cycles were minimal.Conclusions: Doxil®, dexamethasone and low dose thalidomide represents an active induction therapy for patients with untreated multiple myeloma. Major responses were achieved rapidly, arguing for early stem cell harvest and autologous transplant after two cycles of therapy. Treatment related complications occurred early in the course of treatment. Full dose anticoagulation to prevent deep venous thromboembolism is essential when using this combination regimen. No adverse impact on stem cell harvest is associated with this therapy.

Targeted adriamycin delivery to MXT-B2 metastatic mammary carcinoma cells by transferrin liposomes: Effect of adriamycin ADR-to-lipid ratio

In the protein-targeted therapy for cancer, transferrin (Tf) is used to reach a selective and specific target in cancer cells. Tf is used conjugated to chemotherapeutic drugs, insulin, toxins, antibodies, polymers, nanoparticles, lipoplexes and liposomes. Using this latter approach, hydrophobically derivatized Tf was incorporated to liposomal bilayers. The biological activity of Tf-liposome was tested using MXT-B2 cells, a metastatic mammary carcinoma cell line. In Tf binding assays, the Scatchard analysis indicated 4.5x10(5) Tf receptors/cell. In cell growth assays, Tf-liposomes stimulated cell growth in a dose-dependent manner, up to a maximum of 32% of the total free Tf stimulation. Following this, we prepared Tf-liposomes encapsulating adriamycin (ADR) at two different ADR-to-lipid ratios. In vitro cytotoxicity assays against MXT-B2 cells gave IC(50) values 2.1-times lower for Tf-liposomal ADR in comparison to control liposomal ADR. However, similar IC(50) values were found for low ADR-to-lipid ratio Tf-liposomal ADR, as well as for control liposomal ADR. The free Tf added in excess increased the IC(50) value of Tf-liposomal ADR by 51%, while the IC(50) value of control liposomal ADR was unaffected, supporting a receptor-mediated mechanism of targeting by Tf. In addition, the lower IC(50) value is correlated with a higher total of ADR accumulation in the cells.

Applicability of anti-neovascular therapy to drug-resistant tumor: Suppression of drug-resistant P388 tumor growth with neovessel-targeted liposomal adriamycin

Anti-neovascular therapy, one of the effective anti-angiogenic chemotherapy, damages new blood vessels by cytotoxic agents delivered to angiogenic endothelial cells and results in indirect eradication of tumor cells. We previously reported that liposomes-modified with a pentapeptide, Ala-Pro-Arg-Pro-Gly (APRPG-Lip) homing to angiogenic site, highly accumulated in tumor tissue, and APRPG-Lip encapsulating adriamycin (APRPG-LipADM) effectively suppressed tumor growth in tumor-bearing mice. In the present study, we examined the topological distribution of fluorescence-labeled APRPG-LipADM as well as TUNEL-stained cells in an actual tumor specimen obtained from Colon 26 NL-17 carcinoma-bearing mice. The fluorescence-labeled APRPG-Lip dominantly localized to vessel-like structure: A part of which was also stained with anti-CD31 antibody. Furthermore, TUNEL-stained cells were co-localized to the same structure. These data indicated that APRPG-LipADM bound to angiogenic endothelial cells and induced apoptosis of them. We also investigated the applicability of anti-neovascular therapy using APRPG-LipADM to ADM-resistant P388 solid tumor. As a result, APRPG-LipADM significantly suppressed tumor growth in mice bearing the ADM-resistant tumor. These data suggest that APRPG-LipADM is applicable to various kinds of tumor including drug-resistant tumor since it targets angiogenic endothelial cells instead of tumor cells, and eradicates tumor cells through damaging the neovessels.

Effect of small-sized liposomal Adriamycin administered by various routes on a metastatic breast cancer model

The antitumor effects of small-sized liposomal Adriamycin (LADR) administered by various routes were investigated in rabbits bearing well-developed VX2 tumors in the mammary gland. Rabbits received s.c. or i.v., or s.c. combined with i.v., injections of LADR 6 weeks after tumor implantation. The i.v. route showed a significant inhibitory effect on breast tumors and distant metastases. In comparison, metastases in axillary and mediastinal lymph nodes were more efficiently inhibited after s.c. injection. LADR administered concurrently by both the i.v. and s.c. routes produced satisfactory therapeutic activities on both primary breast tumors and metastases in local-regional lymph nodes, lungs and liver, as shown by slowed growth rates, decreased mRNA expression of proliferating cell nuclear antigen, and extensive necrosis and apoptosis of tumor cells. It is concluded that small-sized LADR administered s.c. provides reliable efficacy on lymphatic metastases of breast cancer and that the addition of treatment by the s.c. route to that by the conventional i.v. route can be recommended as a promising procedure to enhance chemotherapeutic effects in patients with metastatic breast cancer.

Lymphatic chemotherapy induces apoptosis in lymph node metastases in a rabbit breast carcinoma model

The purpose of the study was to evaluate the potential of lymphatic chemotherapy in inducing apoptosis in axillary lymph node metastases in a rabbit breast cancer model. A total of 30 female New Zealand rabbits with mammary implantation of VX2 carcinomas were divided into three groups randomly, with ten in each. Treatment was carried out once axillary lymph node reached 5 mm in the maximum diameter. Group A received a subcutaneous injection of liposomal adriamycin (LADR) adjacent to the breast tumor. Group B received free adriamycin (FADR) administered into the auricular vein. Group C received a sham treatment. The dose of adriamycin in each administration was 1 mg/kg in groups A and B. Treatment was repeated every 48 h. Axillary lymph nodes were dissected out 48 h after the third treatment. The nodal sizes before and after the treatment were measured. The therapeutic effect was evaluated in terms of the node volume ratio and apoptotic index (AI) of metastatic cells in nodes identified with TUNEL technique. The significance of difference was determined with one-way ANOVA followed by the Fischer LSD test. Compared to group C, the enlargement of lymph nodes was sufficiently slowed down in both groups A and B, and group A showed a further strong inhibitory effect than group B (P=0.002). Apparent VX2 cell apoptosis was detected in the lymph nodes of groups A and B. The average AI in group B (15.31%) was significantly higher than in group C (5.16%). The highest AI was found in animals of group A (21.73%), with a further significant difference from group B (P=0.000). These data suggest that lymphatic chemotherapy appears to be a promising method to induce apoptosis in lymph node metastases and holds potential in the treatment of advanced breast cancer.

Pentapeptide YIGSR-mediated HT-1080 fibrosarcoma cells targeting of adriamycin encapsulated in sterically stabilized liposomes.

In the peptide-targeted therapy for cancer, peptides are used to reach a selective and specific target in cancer cells. Peptides are used free or coupled to chemotherapeutic drugs, phagues, proteins, polymers, liposomes, and polymer-grafted liposomes. Using this latter approach, the pentapeptide YIGSR was coupled to the distal end from carboxyl groups of liposome-grafted polyethyleneglycol (PEG) chains (YIGSR-PEG-liposome). As a control, the peptide PEAGD coupled to PEG-liposome was used. The biological activity of YIGSR-PEG-liposome was tested using HT-1080 human fibrosarcoma cells. In adhesion assays, the YIGSR-PEG-liposome coated to plastic plates promoted 30% of the specific cell attachment. In competition assays, YIGSR-PEG-liposome inhibited the specific attachment of cells to laminin-1-coated plates by 25%. Following this, we prepared peptide-PEG-liposomes encapsulating adriamycin (ADR). In vitro cytotoxicity assays against HT-1080 cells gave IC(50) values 2.1 times lower for YIGSR-PEG-liposomal ADR in comparison to PEAGD-PEG-liposomal ADR. The free peptide added in excess increased the IC(50) value of YIGSR-PEG-liposomal ADR by 72%, while the IC(50) value of control liposomal ADR was unaffected, supporting a receptor-mediated mechanism of targeting. In addition, the lower IC(50) value is correlated with a higher total of ADR accumulation in the cells.

Anti-neovascular therapy by liposomal DPP-CNDAC targeted to angiogenic vessels

We previously reported that liposomalized 5′- O-dipalmitoylphosphatidyl 2′- C-cyano-2′-deoxy-1-β- D- arabino-pentofuranosylcytosine (DPP-CNDAC), a hydrophobized derivative of the novel antitumor nucleoside CNDAC, is quite useful for cancer therapy. On the other hand, for anti-neovascular therapy, we recently isolated peptides homing to angiogenic vessels from a phage-displayed random peptide library, and observed that peptide-modified liposomal adriamycin strongly suppressed tumor growth, perhaps through damaging angiogenic endothelial cells. In the present study, we modified DPP-CNDAC-liposomes with one of the angiogenic homing peptides, APRPG, and examined their antitumor activity. Three doses of APRPG-modified DPP-CNDAC-liposomes (15 mg/kg as CNDAC) strongly inhibited tumor growth compared with the same number of doses of unmodified DPP-CNDAC-liposomes. The life span was increased 31.8%, with one completely cured mouse out of the six mice treated. Since the accumulation of liposomes in the tumor tissue was not so much different between APRPG-liposomes and non-modified liposomes, the enhanced therapeutic efficacy may be explained as the alteration of targets, i.e. APRPG-modified DPP-CNDAC-liposomes caused tumor growth suppression through damage of angiogenic endothelial cells. Anti-neovascular therapy promises no drug resistance, and should be effective against essentially any kind of solid tumor; and thus the present results demonstrate another benefit of the therapy, namely, high efficacy of cancer treatment.

Antimetastatic effect of hepatotropic liposomal adriamycin on human metastatic liver tumors.

Adriamycin (ADM) was encapsulated in a galactose-conjugated hepatotropic liposome (hLip-ADM) and its ability to enhance the antitumor effect while reducing toxicity was compared with that of free ADM and a control Lip-ADM (cLip-ADM), in two in vitro animal models. Liver metastases were induced in rats by an intravenous injection of 8 x 10(6) AH-130 rat hepatoma tumor cells. All forms of the ADM completely inhibited liver metastasis when given at a dose of 5 mg/kg on day 14 after tumor implantation, whereas liver metastatic foci were observed in six of ten rats in the control group. The reduction in ADM toxicity by liposomalization was remarkable, as significant body weight loss was observed only in the free ADM group, in which four of ten rats died. Additional experiments utilized a human colon cancer xenograft (TK-4) to induce the growth of the liver metastases in mice by orthotopic implantation. The hLip-ADM completely inhibited liver metastasis in rats (0/11), whereas liver metastases developed in 10 of 12 mice in the control group and in 5 of 12 mice given cLip-ADM. Interestingly, liposomal ADM did not have a significant inhibitory effect on transplanted tumor growth assessed 6 weeks after transplantation. These findings indicate that hLip-ADM may be an effective strategy for inhibiting liver metastases from human colon cancer.

Utility of liposomes coated with polysaccharide bearing 1-amino-lactose as targeting chemotherapy for AH66 hepatoma cells.

The cell recognition element is very important for drug delivery systems. We synthesized cholesteryl pullulan (CHP) bearing 1-aminolactose (1-AL) and introduced a saccharide, cholesteryl pullulan bearing 1-aminolactose (1-AL/CHP), to an outer layer of the conventional liposome as a cell recognition element. Lectin recognized the beta-galactose by aggregation of 1-AL/CHP coated liposome (1-AL/CHP liposome). The uptake of this liposome to AH66 rat hepatoma cells was greater than in liposomes without 1-aminolactose in vitro. Furthermore, 1-AL/CHP liposomal adriamycin showed a stronger antitumor effect in comparison with other types of liposomal adriamycin in vitro. When in vivo tumor-targeting efficacy was investigated in AH66 tumor transplanted mice using 3H-liposome, the tumor/serum radioactivity ratio in mice injected with 1-AL/CHP liposome was higher than that of mice injected with other liposomes. These observations suggest that 1-AL is effective as a cell recognition element. As a result, 1-AL/CHP liposome is considered to be a good carrier of anticancer drugs for the active targeting of tumor cells.

Ｐｏｌｙｅｔｈｙｌｅｎｅｇｌｙｃｏｌ修飾ａｄｒｉａｍｙｃｉｎ含有リポソームの抗腫よう効果および副作用

We examined the effects of polyethyleneglycol (PEG)-coated liposome containing adriamycin (PEG-LADR)on the antitumor activity and toxic side effect of adriamycin (ADR), using Ehrlich ascites carcinoma bearing CDF1, male mice. PEG-LADR significantly enhanced the antitumor activity of ADR against the solid tumor. This increment of antitumor activity induced by PEG-coated liposomalization supported by the elevation of ADR concentration in the tumor. Next, we examined two biochemical parameters, lipid peroxide (LPO) level and glutathione peroxidase (GSHpx) activity, as indices of ADR induced side effects. There were the increase of LPO level and the decrease of GSHpx activity in the heart of mice after ADR solution administration. ADR concentration in the heart on the 24th hour after PEG-LADR administration showed higher than that after ADR solution treatment. However, PEG-LADR administration were not changed LPO level and GSHpx activity in the heart, compared to normal level. In the liver and the pancreas, same results were shown. Furthermore, regarding the lethal effect of ADR, the acute toxicity after PEG-LADR treatment significantly reduced, compared to that after ADR solution treatment. Therefore, these results suggested that PEG-LADR can significantly increase the antitumor activity of ADR solution with decreasing its side effects, and the side effects of liposomal ADR should not evaluated by ADR concentration in each tissues.

Endocytosis does not necessarily augment the cytotoxicity of adriamycin encapsulated in immunoliposomes

We studied the relationship between endocytosis and cytotoxicity of adriamycin (ADM) encapsulated in antibody-coated liposomes (immunoliposomes, IL) which are called chemoimmunoliposomes (CIL), by using several human cancer cell lines. IL coated with a monoclonal antibody, HBJ127 (IgG), which recognizes human gp125 antigen, specifically bound to gp125-positive target cancer cell lines, KU-1, T24, MKN-7, SKBr-3 and LS174T. Flow cytometric analysis using IL encapsulating carboxyfluorescein (CF) revealed that efficiencies of endocytosis varied among different cancer cells. The rate of IL internalization was in the order KU-1> T24> MKN-7> SKBr-3> LS174 T . In 1 h incubation at 37°C, all the four cell lines other than LS174T internalized about 60% of IL which were bound on their cell surfaces. KU-1, T24 and MKN-7, but not SKBr-3, significantly processed IL in endosome or lysosome. On the contrary, 80% of CIL bound to LS174T remained on the cell surface even after 2 h incubation. Furthermore, we evaluated the cytotoxic activities of CIL against the same panels of cancer cells. CIL inhibited the growth of all cancer cells tested in antibody-dependent manner, but, contrary to our expectation, KU-1 and T24 cells, which showed significant endocytosis activity, required a 7–14-fold higher amount of ADM binding than LS174T cells with low endocytosis activity for 50% cell growth inhibition. The difference of sensitivity to free ADM was only within 2.3-fold among those cancer cells. These results showing that liposomal ADM endocytosed is less effective than that remaining on the cell surface suggest that endocytosis is not necessarily required for cytotoxicity of CIL.