Mustard Group Research Articles

Gösta Jönsson (1909-1978): A pioneer in the hormonal treatment of prostate cancer in Sweden.

Treatment of castration-resistant prostate cancer is and has been a challenge. In 1957, the chemist Imre Könyves came to Sweden as a refugee from Hungary and started to work at AB Leo, a pharmaceutical company in Helsingborg. In 1961, he started to synthesize compounds where the oestrogens were linked to a mustard group by a carbamate. This resulted in estramustine phosphate, which was initially tested against mammary cancer with disappointing results. He then started a cooperation with urology professor Gösta Jönsson, Head of the Department of Urology at the Lund University Hospital, to test estramustine phosphate against prostate cancer. Jönsson started clinical estramustine phosphate tests in 1966. His studies were one-armed and consecutive, with a "favourable response" in 83% of previously untreated patients. These favourable results could not be reproduced in later randomized controlled studies suggesting that estramustine phosphate as primary treatment was not better than conventional estrogenic treatment. Conclusions: Even if the results of Gösta Jönsson's studies could not be confirmed, the subsequent randomized studies of estramustine phosphate may hide the desired action of estramustine phosphate in a subgroup of patients. It has still not been elucidated whether estramustine phosphate has effects in this subgroup of patients with ostrogen-resistant prostate cancer.

Genetic variability, traits interactions, and principal component analysis of broad-leaf mustard group (Brassica juncea L.) to focus on the yield improvement

Mustard (Brassica spp.), one of the most important oilseed crops, is currently the third-largest source of vegetable oil in the world. In Rabi, 2022–2023 at the Regional Agricultural Research Station (RARS), Barishal, research of Brassica juncea L. was carried out with 10 genotypes to assess yield and yield contributing features. The RCBD design for the experiment included three replications. Plant height, the number of primary branches, the number of secondary branches, and the number of seeds per siliqua were the characteristics for which the analysis of variance findings indicated that there were very significant differences. Using the UPGMA cluster analysis, four distinct clusters were found among the analyzed accessions. Cluster II obtained the most accessions (7), averaging 1765.28 kgha-1, representing 25.20% of the average group mean yield. A correlation study focuses on the highly significant and positive correlation between a crop’s associated features and its yield. The overall amount of variance was found to be 46.18% for PC1 and 17.61% for PC2 using Principal Component Analysis (PCA). Maturity duration ranged from 106-110 days while the 50% flowering period varied from 43-46 days. Seed yield ranged from 1463 to 1970 kgha-1 and BJ-11536(12)-1 produced the highest yield among the genotypes. Which was it produced 23.58% and 20.41% higher yields than check varieties BARI Sarisha-11 and BARI Sarisha-19, respectively. The second highest yield was recorded from BJDH-05 which produced 18.90% and 15.85% higher yield over the check varieties BARI Sarisha-11 and BARI Sarisha-19, respectively. The lowest seed yield (1463 kgha-1) was obtained from the entry BJ-53611(12)-8. Based on, statistical analysis considering seed yield and other yield contributing characters, four lines BJ-11536(12)-1, BJDH-05, BJDH-20, BJ-11536(12)-5, and BJ-1110(12)-1 were noted as high-yielding accessions and could be used for future breeding program.

An oxygen-carrying and lysosome-targeting BODIPY derivative for NIR bioimaging and enhanced multimodal therapy against hypoxic tumors.

Cancer treatment modalities have gradually shifted from monotherapies to multimodal therapies. It is still a challenge to develop a synergistic chemo-phototherapy system with relieving tumor hypoxia, specific targeting, and real-time fluorescence tracking. In this study, we designed a multifunctional BODIPY derivative, FBD-M, for synergistic chemo-phototherapy against hypoxic tumors. FBD-M was composed of four parts: 1) The BODIPY fluorophore selected as a theranostic core, 2) A pentafluorobenzene group modified on meso-BODIPY to carry oxygen, 3) A morpholine group hooked to one side of BODIPY served as a lysosome-targeting unit for enhancing antitumor effect, and 4) An aromatic nitrogen mustard group introduced on other side of BODIPY to achieve chemotherapy. After introducing the morpholine and aromatic nitrogen mustard in BODIPY, the conjugate system of BODIPY was also expanded to realize near-infrared (NIR) phototherapy. Finally, FBD-M was obtained by a rational design, which possessed with NIR absorbance and emission, photosensitive activity, oxygen-carrying capability for relieving tumor hypoxia, high photothermal conversion efficiency, good photostability, lysosome targeting, low toxicity, and synergistic chemo-phototherapy against hypoxic tumors. FBD-M had been successfully applied for anticancer in vitro and in vivo. Our study demonstrates that FBD-M can serve as an ideal multifunctional theranostic agents.

Evaluation of Fungicides and Plant extracts against stem rot of mustard pathogen (Sclerotinia sclerotiorum) In-Vitro and In-Vivo condition

Mustard group of crops are important oil seed crops of the world. The stem rot of mustard incited by Sclerotinia scletiorum (Lib.) de Bery, is a new emerging drastic disease. The pathogen was tested with 12 Fungicides and 03 Plant extract In-Vitro.In-Vivo condition 9 fungicides and 01 plant extract were tested in the year of 2019-20 and 2020-21. Among the tested fungicides namely Mancozeb, Chlorothalonil, and Vitvax were found most effective which inhibited the growth of the pathogen completely and inhibition exhibited 100%. Vapum was the least effective fungicide which showed 88.03mm fungal growth with 2.18 per cent inhibition over control. In-Vivo condition Chlorothalonil and Mancozeb was most effective fungicides which was showed mean minimum disease incidence (06.25% and 08.87%) and maximum mean yield 6.55 and 5.75q/ha. respectively. Next order of superiorly fungicides were Kitazin, Bavisin and Vitavax, which were showed the mean disease incidence 11.68, 13.65, 15.6 and yield 5.20, 4.70 and 4.30q/ha. respectively. Among the tested fungicide Zineb was least effective which was showed the maximum mean incidence of disease (30.03) with respect to minimum mean yield (2.67q/ha). Among the plant extract Nimbidinwas least effective which showed 33.14 per cent mean disease incidence and minimum (2.32q/ha) grain yield.

Biomarker sST2 in Adults with Transposition of the Great Arteries Palliated by Mustard Procedure: A Five-Year Follow-up.

The Mustard procedure was an early cardiac surgery for transposition of the great arteries (TGA). Despite being successful, it has been associated with long-term arrhythmias and heart failure. A key factor complicating management in adults with congenital heart disease (CHD) is the deficiency of biomarkers predicting outcome. Soluble suppression of tumorogenicity-2 (sST2) is secreted by cardiomyocytes in response to mechanical strain and fibrosis. We hypothesized that adults with a Mustard procedure would have higher levels of sST2 than healthy individuals, and this would correlate with clinical outcome. We performed a single-center study in patients managed during childhood with a Mustard procedure versus age-matched controls. Clinical and demographic data were collected and biomarkers (sST2, cTnI, BNP, lipid panel, insulin, and glucose) were obtained. There were 18 patients (12 male) in the Mustard cohort and 18 patients (6 male) in the control group (22-49years, mean of 35.8 vs. mean 32.6years, respectively, p = ns). Nine Mustard subjects were NYHA class II, and 9 subjects were class III. The control group was asymptomatic. sST2 in the Mustard group was elevated in 56% vs. 17% in controls (p = 0.035). Of the Mustard subjects with elevated sST2, 60% had elevated cTnI and BNP, and 90% had low HDL. Over five years, the Mustard patients with elevated sST2 values had greater medication use, arrhythmias, hospitalizations, and ablation/pacer implantations than Mustard subjects with normal sST2. Mustard subjects with elevated sST2 had other biomarker abnormalities and clinically worse outcomes. Thus, sST2 may add a predictive value to cardiac-related morbidity and mortality.

Optimization of Intra-Row Spacing for Yield Enhancement in System of Mustard Intensification (SMI) Techniques

The experimental work was carried out during the rabi seasons (November to February) of the year 2018 and 2019 at Instructional field of RRS (OAZ), Uttar Banga Krishi Viswavidyalaya, Majhian, Dakshin Dinajpur, West Bengal, India to study the growth and yield performance of different varieties from toria, rapeseed and Indian mustard with three different spacing combinations under System of Mustard Intensification (SMI) technology. The seedlings were raised in micro-pots (2×2.5 cm2 size) and transplanted in the main plot. The experiment was laid out in a completely randomized block design replicated thrice. The treatments consisted of three levels of plant spacing (25×25 cm2, 45×45 cm2, 60×60 cm2) and included 3 varieties one each from toria, rapeseed and Indian mustard group. Among different treatment combinations, variety Bhagirathi (from Indian mustard group), in combination with spacing of 60×60 cm2 (S3V3) was recorded with highest yield (1348.08 kg ha-1) followed by its rest of the combinations with 25×25 cm2 (1331.42 kg ha-1) and 45cm×45 cm (1320.67 kg ha-1), i.e. S1V3 and S2V3, respectively. Plant height (cm) was also recorded highest with S3V3 treatment followed by S2V3 and S1V3. Total chlorophyll (mg 100 g-1) content was recorded highest with the same Bhagirathi variety S1V3 treatment (18.99) followed by S2V3 (17.45) and S3V3 (17.61). Amongst the yield attributing characters, number of secondary branches, number of siliqua plant-1 and total chlorophyll content were recorded highest with the Bhagirathi variety from the Indian mustard group with the spacing of 60×60 cm2 (S3V3).

Biomarker sST2 in Adults with Transposition of the Great Arteries Palliated by Mustard Procedure

Background: Transposition of the great arteries (TGA) is a congenital heart defect (CHD) in which the aorta and pulmonary artery are transposed; it requires urgent surgical intervention. The Mustard procedure was an early surgery allowing survival into adulthood. However, this procedure is associated with long-term adverse effects including arrhythmias and heart failure (HF). A key factor impairing HF management in adults with CHD is lack of biomarkers to predict outcome. Soluble ST2 (sST2) is a protein secreted by myocytes in response to mechanical strain and fibrosis. No studies have focused specifically on sST2 in TGA individuals that underwent a Mustard procedure. In this study, we hypothesized adults with TGA palliated with a Mustard procedure would have higher levels of sST2 than healthy individuals, and this would correlate with functional class status. 
 
 
 Methods: We screened the Pediatric Cardiology database for D-TGA subjects and Mustard operation. Healthy controls were recruited from clinic and the community. Patients were matched by age to a control group of 21 healthy individuals. Severity of symptoms was assessed by NYHA functional classification. sST2 levels were obtained using Critical Diagnostics Presage ST2 Assay kit. Additionally, cTnI, BNP, lipid panel, insulin, glucose, and EKG and echocardiography (TGA) were obtained at IUSM clinical laboratories. 
 
 Results: We identified 45 patients with D-TGA and Mustard operation. 19 patients were included in analysis: 12 male and 7 female subjects aged 18 to 46, mean of 34.9 years. In the Mustard group, 9 subjects were assigned to NYHA class II, and 9 to class III. The control group was asymptomatic. sST2 levels in the Mustard group were elevated in 53%, while only 29% in the controls. Of the Mustard subjects with elevated sST2, 67% had elevated cTnI, 80% arrhythmias, 70% EKG T-wave abnormalities, and 80% took cardiac medications. 90% of these patients had low HDL, 40% high LDL, and 40% low insulin. 
 
 Conclusion: This study demonstrates patients with Mustard operation averaged higher sST2 levels than healthy subjects. Importantly, of the patients with elevated levels, there was a significant association of sST2 with biomarker abnormalities and clinical heart failure signs. This suggests in patients with D-TGA palliated with Mustard procedure, sST2 may add predictive value to cardiac related morbidity and mortality.

MP-Pt(IV): A MAOB-Sensitive Mitochondrial-Specific Prodrug for Treating Glioblastoma.

We have previously reported the in vitro and in vivo efficacy of N,N-bis(2-chloroethyl)-2-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)propenamide (MP-MUS), a prodrug that targeted the mitochondria of glioblastoma (GBM). The mitochondrial enzyme, monoamine oxidase B (MAOB), is highly expressed in GBM and oxidizes an uncharged methyl-tetrahydropyridine (MP-) moiety into the mitochondrially targeted cationic form, methyl-pyridinium (P+-). Coupling this MAOB-sensitive group to a nitrogen mustard produced a prodrug that damaged GBM mitochondria and killed GBM cells. Unfortunately, the intrinsic reactivity of the nitrogen mustard group and low solubility of MP-MUS precluded clinical development. In our second-generation prodrug, MP-Pt(IV), we coupled the MP group to an unreactive cisplatin precursor. The enzymatic conversion of MP-Pt(IV) to P+-Pt(IV) was tested using recombinant human MAOA and rhMAOB. The generation of cisplatin from Pt(IV) by ascorbate was studied optically and using mass spectroscopy. Efficacy toward primary GBM cells and tumors was studied in vitro and in an intracranial patient-derived xenograft mice GBM model. Our studies demonstrate that MP-Pt(IV) is selectively activated by MAOB. MP-Pt(IV) is highly toxic toward GBM cells in vitro MP-Pt(IV) toxicity against GBM is potentiated by elevating mitochondrial ascorbate and can be arrested by MAOB inhibition. In in vitro studies, sublethal MP-Pt(IV) doses elevated mitochondrial MAOB levels in surviving GBM cells. MP-Pt(IV) is a potent chemotherapeutic in intracranial patient-derived xenograft mouse models of primary GBM and potentiates both temozolomide and temozolomide-chemoradiation therapies. MP-Pt(IV) was well tolerated and is highly effective against GBM in both in vitro and in vivo models.

Autophagy Inhibition Potentiates the Anticancer Effects of a Bendamustine Derivative NL-101 in Acute T Lymphocytic Leukemia.

Acute T lymphocytic leukemia (T-ALL) is an aggressive hematologic resulting from the malignant transformation of T-cell progenitors. Drug resistance and relapse are major difficulties in the treatment of T-ALL. Here, we report the antitumor potency of NL-101, a compound that combines the nitrogen mustard group of bendamustine with the hydroxamic acid group of vorinostat. We found NL-101 exhibited efficient antiproliferative activity in T-ALL cell lines (IC50 1.59–1.89 μM), accompanied by cell cycle arrest and apoptosis, as evidenced by the increased expression of Cyclin E1, CDK2, and CDK4 proteins and cleavage of PARP. In addition, this bendamustine-derived drug showed both a HDACi effect as demonstrated by histone hyperacetylation and p21 transcription and a DNA-damaging effect as shown by an increase in γ-H2AX. Intriguingly, we found that NL-101-induced autophagy in T-ALL cells through inhibiting Akt-mTOR signaling pathway, as indicated by an increase in LC3-I to LC3-II conversion and decrease of p62. Furthermore, inhibition of autophagy by 3-methyladenine increased apoptotic cell death by NL-101, suggesting a prosurvival role of autophagy. In summary, our finding provides rationale for investigation of NL-101 as a DNA/HDAC dual targeting drug in T-ALL, either as a single agent or in combination with autophagy inhibitors.

Discovery of N-(2-Aminophenyl)-4-(bis(2-chloroethyl)amino)Benzamide as a Potent Histone Deacetylase Inhibitor.

Inhibition of histone deacetylases (HDACs) has been an important emerging therapy for the treatment of multiple cancers. However, the application of HDAC inhibitors is restricted by the limited potency against solid tumors. In order to discover novel HDAC inhibitors with potent antitumor activities, nitrogen mustard group was introduced to the structure of CI994. The derived molecule N-(2-aminophenyl)-4-(bis(2-chloroethyl)amino)benzamide (NA) exhibited enzyme inhibitory pattern of class I selectivity with IC50 values of 95.2, 260.7, and 255.7 nM against HDAC1, HDAC2, and HDAC3, respectively. In the antiproliferative assay, NA exhibited 10.3-fold (2.66 μM) and 11.3-fold (1.73 μM) higher potency than did suberoylanilide hydroxamic acid (SAHA) (27.3 and 19.5 μM) in inhibition of A2780 and HepG2 cell growth, respectively. Further HepG2 cell-based cell cycle and apoptosis studies revealed that induction of the G2/M phase arrest and cell apoptosis contributes to the antitumor effects of NA. It is suggested that NA could be utilized as a lead compound in the development of bifunctional HDAC inhibitors for the treatment of solid tumors.

Investigation on the cytotoxic effects of nitrogen-mustard-derived Schiff bases. Studies on the reactivity of the N-mustard pharmacophoric group

4-[bis(2-chloroethyl)amino]benzaldehyde-N(4)-phenyl-thiosemicarbazone (1), 4-[bis(2-chloroethyl)amino]benzaldehyde-N(4)-para-chloro-phenyl-thiosemicarbazone (2), 4-[bis(2-chloroethyl)amino]benzaldehyde-N(4)-methyl-thiosemicarbazone (3) and 4-[bis(2-chloroethyl)amino]benzaldehyde-benzoylhydrazone (4) were obtained by functionalization of the bis(2-chloroethyl)amino DNA alkylating group with a thiosemicarbazone/hydrazone moiety. The compounds were fully characterized by means of their infrared (including theoretical vibrational analyses) and NMR spectra, as well as by means of X-ray crystal structure determinations.When assayed on a panel of different tumor cells only (1) exhibited cytotoxic effects on B16F10 melanoma cells (IC50 = 17.1 ± 3.1 μM). A cell viability assay on melanoma cells revealed that while (1) had cytotoxic effects on B16F10 and A2058 cells, the structure-related hydrazone (4) was devoid of activity. In addition, (1) was capable to reduce viability of melanoma cells without affecting the viability of non-malignant human endothelial cells (HUVEC), suggesting that this compound or its analogues could be further investigated as potential antimelanoma drug candidates. Preliminary investigation on the mode of action of (1) indicated that the compound induces cell death by necrosis.Theoretical calculations showed that in all compounds (1–4) the reactivities of the N-mustard pharmacophoric group towards dehalogenation are similar. In addition, theoretical studies suggested that the nucleophilicities of the mustard group are very similar in (1–4). Hence, all compounds (1–4) might be expected to exhibit comparable reactivity, and the differences in biological activity among the compounds may not be attributed to variances in reactivity at the N-mustard group.

Mustard group chemical warfare agents and clinical effects

Mustard gazi gectigimiz yuzyil icerisinde en cok kullanilan, yuz binlerce insanin olumune ve yaralanmasina yol acan vezikan ajanlar olarak siniflandirilan kimyasal silah grubudur. Sulfur mustard (SM) ve analogu olan Nitrojen mustard (NM)bu gruba dahil iki kimyasal ajandir. Icinde bulundugumuz orta dogu ve yakin cografyadaki Iran, Irak, Libya, Suriye gibi ulkelerin elinde tonlarca sulfur mustard gazi bulundugu ve bunun bolge ulkeleri icin cok ciddi bir tehdit oldugu artik herkes tarafindan bilinen bir gercektir. Ortaya cikardigi devasa etkiler nedeniyle mustard gazi “savas gazlarinin krali”unvaninialmistir. Tarih surecinde ilk defa 1.Dunya Savasi’nda kullanilmis olan mustard gazi, takip eden yillar icerisinde defalarca silahli mucadelelerde kullanilmis asker ve sivillerin kitlesel olumune ve yaralanmasina neden olmustur. Elde edilmesi, depolanmasi ve kullanimi konusundaki kolaylik mustard gazinin tercih edilmesinin en onemli nedenleridir. Mortaliteden cok mobiditeye neden olmasimustard gazinin bir savas ajani olarak tercih edilmesinin diger nedenidir. Bu nedenle devletler yillarca yaralilarin bakimiyla ugrasmis ve bu durum maddi ve manevi bir yikima neden olmustur. Oldukca yuksek toksisiteye sahip olmasi maruz kalinan doz ile dogru orantili olarak ani olumleri de beraberinde getirmektedir. Baslica etkilerini goz, akciger ve deri uzerinde gosteren mustard gazi kanser olusumu da dahil olmak uzere vucutta bircok hasara yol acmaktadir. Nitrojen mustard kimyasal yapisi ve olusturmus oldugu toksisite bakimindan SM ile buyuk olcude benzerlik gostermektedir. Olusturmus oldugu etkiler nedeniyle kullanimi yasaklanmistir. Kullanimi sonrasinda yaralanan kisiler uzerinde yapilan calismalar kemoterapotik bir ajan olarak kullanilabilecegi sonucunu dogurmustur. Antidotu bulunmayan tek kimyasal ajan olmasi nedeniyle yaklasik yarim yuzyildir uzerinde yurutulen antidot gelistirme cabalari belli bir duzeye kadar gelmistir ancak bu konuda halen alinmasi gereken onemli mesafeler bulunmaktadir

Effect of CP Metronomic Chemotherapy on RPMI 8226 Cell Proli-feration and Notch1/NF-κB Signaling Pathway In Vitro

To investigate the effect of metronomic chemotherapy of low dose phosphoramide combined with prednisolone (CP metronomic chemotherapy) on proliferation and apoptosis of RPMI 8226 cells, and to explore its regulating effect on Notch1/NF-κB signaling pathways. Experiment was divided into the DMSO control group, and the phosphoramide mustard (PM) group, the prednisolone group, the phosphoramide mustard plus prednisolone group (the CP group). RPMI 8226 cells were treated with different drugs, CCK-8 method was used to detect cell proliferation, flow cytometry was used to detect the cell cycle and apoptosis, reverse transcription PCR was used to detect Notch1 and NF-κB mRNA expression level. Compared with DMSO control group, RPMI8226 cell proliferation inhibition rate in all the PM, prednisolone and CP groups increased significantly with prolonging of time (r of 0.994,0.996,0.999, respectively, P<0.001). And at the same time, the inhibitory rate of cell proliferation was significantly different; the cell inhibitory rate in PM group was lowest, that in CP group was highgest, that in prednissone group was intermediate (P<0.01). After 48 hours, compared with the DMSO control group, the G1/G0 cell proportion in treatment group increased significantly, S phase cell proportion decreased significantly, especially in PM and CP groups. The G2/M phase cell proportion increased in PM group, while reduced in the prednisolone and the CP groups. After 48 hours, compared with the DMSO control group, RPMI 8226 cell apoptosis rate increased as follow: in PM, pre-dnisolone and CP group(P<0.01). After 48 hours, compared with the DMSO control group, Notch1 and NF-κB mRNA expression in the prednisolone, the PM and the CP group decreased significantly(P<0.001). CP metronomic chemotherapy can significantly reduce RPMI 8226 cell proliferation, promote RPMI 8226 cell apoptosis, arrest RPMI 8226 cells mainly in the G1/G0 phase, and significantly reduce Notch1 and NF-κB expression levels. It is suggested that Notch1/NF-κB signaling pathways is involved in CP metronomic chemotherapy for MM.

Synthesis, cytotoxicity, topoisomerase I inhibition and molecular docking of novel phosphoramide mustard sophoridinic acid analogues

A series of novel phosphoramide mustard sophoridinic acid analogues, consisting of nitrogen mustard group and sophoridinic acid scaffold, have been designed, synthesized and evaluated for their topoisomerase inhibitory activity as well as cytotoxicity against six tumor cell lines (SMMC‐7721, LoVo, MCF‐7, K562, S180 and H22) and a normal cell line (L929). Among the compounds tested, five were found to be potent inhibitors and exhibited potent cytotoxicity against S180 and H22 cell lines with IC50 values of 1–4 μM. Further mechanistic studies showed that this class of compounds acted as novel topoisomerase I (Topo I) catalytic inhibitors by preventing the binding of Topo I to DNA and inhibiting the cleavage of DNA, and molecular docking studies revealed that the binding energy for these compounds was comparable to that for classic Topo I inhibitors CPT and HCPT, indicating that the compounds have an interaction with DNA and Topo I.

Synthesis, structure-activity relationship and biological evaluation of novel nitrogen mustard sophoridinic acid derivatives as potential anticancer agents.

A series of novel nitrogen mustard sophoridinic acid derivatives were designed, synthesized and evaluated for their cytotoxicity. Of the newly synthesized compounds, compound 6 exhibited a potent effect against hepatocellular carcinoma in vitro and in vivo. SAR analysis indicated that introduction of a nitrogen mustard group to the structure of sophoridinic acid significantly enhance the antitumor activity. Moreover, molecular docking study exhibited benzyl group introduced to the nitrogen atom at the 12-position and aryl nitrogen mustard group at the 4'-carboxyl region for compound 6 were beneficial for the higher anticancer activity. This work provides useful information for further structural modifications of these compounds and for the synthesis of new, potent antitumor agents.

Bendamustine, Bortezomib and Dexamethasone (BVD): A Combination With a Substantial Activity and a Manageable Toxicity In Patients With Relapsed-Refractory Multiple Myeloma (MM)

Bendamustine, a bi-functional alkylating agent comprising a purine-like benzimidazole ring an a nitrogen mustard group, exerts a peculiar mechanism of action if compared to most conventional alkylators and this may partially explain its effectiveness in alkylator-resistant cells. In patients with MM several studies demonstrated the efficacy and tolerability of bendamustine as monotherapy or in combination with new drugs, particularly bortezomib that was found to enhance the in vitro sensitivity of MM cells to bendamustine.These observations represented the rationale for our protocol design namely to evaluate the combination bendamustine (70 mg/m2 days 1, 8), bortezomib (1.3 mg/ m2 days 1, 4, 8, 11) and dexamethasone (20 mg days 1-2, 4-5, 8-9, 11-12) (BVD). Cycles were administered every 4 weeks up to four cycles. Patients achieving a response less than a PR were taken off-study. Patients obtaining at least a PR received two additional treatment cycles followed by a 12-months consolidation phase with cycles repeated every 2 months. Therefore, patients with a PR after the induction phase could receive up to 18 months of treatment and up to 12 cycles of BVD. Patients with relapsed/refractory MM of any age, with adequate cardiac, liver and hematological function, not refractory to bortezomib and treated with no more than four previous lines of therapy were enrolled in this prospective, single-arm, open-label, phase II study conducted in 21 Italian centres. The primary endpoint was achievement of a response at least PR after four cycles of BVD and response was assessed according to to IMWG criteria.From March 2011 to June 2012, 75 patients were included. Median age was 68 years (range 41-85), 26.5% had ISS stage 3, 19% IgA myeloma and 9% renal failure. In total, 8 of 36 evaluable patients (22%) had adverse cytogenetics. All patients had received prior treatment with new drugs, including targeted agents such as thalidomide (57%), lenalidomide (54.5%) or bortezomib (46.5%). Patients had received a median of one prior line of therapy (range 1-4), including alkylators (69%), anthracyclines (29%) and ASCT (44%). Twenty-four patients (32%) were refractory to IMIDs.The response rate ≥ PR after four cycles of BVD was 71.5%, including 11 patients with CR (16%), 13 VGPRs (18.5%) and 26 PRs (37%). Also, 14 patients (20%) had disease stabilization while 6 (8.5%) had progressive disease. Median time to response was 1.2 months (range 0.9-1.4 months). Only prior treatment with bortezomib significantly reduced the response rate ≥ PR (54.5% vs 86.5%; P = 0.003).At a median follow-up of 12 months (range 6-24), 30 patients had progressed and 18 had died. Median TTP and PFS were 16.5 months and 15.5 months, respectively while median OS had not been reached and 78% of patients were alive at 1 year. The Cox regression analysis identified prior therapy with bortezomib plus lenalidomide as the only factor that significantly reduced TTP (9 vs 17 months; HR = 4.5; 95% CI = 1.7-12.3; P = 0.005)Grade 3-4 adverse events occurred in 55% of patients leading to therapy reduction in 20% and to protocol discontinuation in 10.5% of patients. The most frequent severe adverse events were thrombocytopenia (30.5%), neutropenia (18.5%), infections (12%), peripheral neuropathy (8%), gastrointestinal and cardiovascular events (both 6.5%). Compared with younger, patients aged > 70 years had a significantly higher incidence of grade 3-4 thrombocytopenia (22% vs 37%; p=0.042) and severe infections (7 vs 19%; p=0.047) and consequently a higher rate of therapy reduction (9% vs 34.5%; p=0.007) and therapy discontinuation (7% vs 15.5%; p=0.043). Moreover, 4/5 early deaths occurred in patients aged more than 70 years.BVD combination is an effective regimen in relapsed-refractory MM patients since it elicits rapid, high (> 70%) and good quality of response (more than a third of patients achieved CR + VGPR). Moreover, BVD combination is a feasible and well tolerated regimen provided that adapted therapy and adequate antibiotic prophylaxis are employed in patients older than 70 years. Disclosures:Offidani:Mundipharma: Honoraria, Research Funding. Off Label Use: Bendamustine.

Comparison of Late Results of Arterial Switch Versus Atrial Switch (Mustard Procedure) Operation for Transposition of the Great Arteries

Two decades after surgery for transposition of the great arteries, the clinical status, cardiac function, cardiorespiratory performance, and neurohormonal activity of patients who underwent either atrial switch (Mustard) operations or arterial switch operations (ASOs) were compared. Sixty-two patients with simple transposition of the great arteries who underwent either Mustard (n= 34) or ASO (n= 28) procedures were included in this cross-sectional study. Following the same study protocol, clinical workup including echocardiography, stress testing, and blood work was completed for all patients. Mean ages in the 2 groups were comparable, at 20.6 ± 2.1 and 20.6 ± 3.4 years in the ASO and Mustard groups, respectively. All ASO patients were in New York Heart Association class I, whereas 59% of Mustard patients were in class II or III. Peak oxygen uptake was higher in ASO patients (percentage of predicted 80% vs 69%, p <0.01). Compared with healthy subjects, the mean Tei index for systemic ventricle was high in the 2 groups, but this parameter was significantly higher in Mustard than ASO patients (0.60 ± 0.16 vs 0.47 ± 0.14, p <0.01). The median plasma N-terminal pro-brain natriuretic peptide level in ASO patients was within the normal range, but the Mustard group had significantly higher levels (42 ng/ml [range 18 to 323] vs 172 ng/ml [range 26 to 1,018], p <0.0001). In conclusion, this cross-sectional assessment 2 decades after surgery reveals better clinical status in patients who underwent ASO compared with Mustard patients. This holds in terms of cardiac function, cardiorespiratory performance, and neurohormonal activity.

A phase II study of second-line bendamustine in relapsed or refractory small cell lung cancer (SCLC).

7094 Background: Bendamustine is an alkylating agent with a nitrogen mustard group and purine-like benzimidazole group. Bendamustine in combination with carboplatin has shown efficacy as first line therapy in extensive stage SCLC with RR 73%, TTP 5.2 months and OS 8.3 months [Köster, et al, JCO 2007]. This study aims to investigate the efficacy and safety of single agent bendamustine as 2nd or 3rd line therapy in patients with extensive-stage disease, small-cell lung cancer (ED-SCLC). Methods: This is an open-label, single-arm, multicenter phase II trial. Eligible patients had previously treated ED-SCLC, up to 2 prior regimens, ECOG performance status 0-2, evaluable/measurable disease, and adequate marrow, renal and hepatic function. Patients with stable treated brain metastases were allowed. Patients were treated with bendamustine (120mg/m2 IV days 1 and 2 every 3 weeks) for up to 6 cycles. Evaluation occurred every 2 cycles. Primary endpoint was TTP; secondary endpoints include RR, PFS, OS, and toxicity. Results: 48 patients were enrolled; 56% were male and 96% were Caucasian. 33 patients were evaluable for response. There was 1 CR, 9 PR, 13 SD (48% disease control rate) and 10 PD. Median TTP was 3.37 months (95% CI 2.30 to 4.47 months). At the time of analysis, 13 patients were alive and with a median overall survival of 4.77 months (95% CI 3.67 to 6.07 months). 5 patients (10.4%) required dose reductions due to AEs, 2 due to fatigue, 1 due to neutropenia, 1 due to pancytopenia and 1 due to pneumonia. Grade 3/4 AEs included fatigue (18.8%), dyspnea (14.5%), infection without neutropenia (12.5%), anemia (8.3%), neutropenia (8.3%), and diarrhea (8.3%). Conclusions: These data indicate that single agent bendamustine appears to be well tolerated and effective in the second or third line setting for patients with SCLC.

Bendamustine with or without rituximab for the treatment of heavily pretreated non-Hodgkin’s lymphoma patients

Bendamustine is an alkylating agent with a nitrogen mustard group and a purine-like benzimidazole group. The aim of this study was to collect all the Italian experiences with this drug in order to evaluate the results in term of response to therapy and toxicities. We analyzed lymphoma patients treated in 24 Italian haematological centres with bendamustine alone or in combination with anti-CD20 antibody. One hundred seventy-five relapsed or refractory lymphoma patients were enrolled. The median age was 69 years (range 26-87). Seventy-nine patients were relapsed, 35 were refractory and 61 presented a progressive disease after partial response. The diagnoses were 60 indolent non-follicular lymphomas, 34 diffuse large B-cell lymphomas, 48 follicular lymphomas, 30 mantle cell lymphomas and three peripheral T-cell lymphomas. All patients were evaluable for response: 52 (29%) with complete remission, 72 (43%) with partial response with an overall response rate of 71%, and 51 non-responders. With a median observation period of 10 months (1-43), 70% of patients are alive. In summary, this retrospective study shows that treatment with bendamustine alone or in combination with rituximab is a safe and effective regimen in a subset of multi-resistant patients.

Safety, Tolerability and Activity of Ofatumumab, Bendamustine and Dexamethasone Combination As First-Line Treatment of Mantle-Cell Lymphoma in the Elderly: A Multicenter Study

Abstract 1647 Background and Aims:Mantle cell lymphoma (MCL) is the aggressive subtype of non Hodgkin’s lymphoma with the poorest long-term survival probability. There is no consensus for its treatment: in most cases, R-CHOP immuno-chemotherapy, with its 34% complete remission (CR) rate, is still considered the standard of care. Very encouraging results have been reported using more aggressive induction treatments as first-line therapy but these have a role only in the initial management of clinically fit patients less than 60–65 years of age. Since the median age at diagnosis is 63 years and intensive treatment tend to be poorly tolerated by elderly patients, a frontline therapy that can be administered to this category of patients is needed. The main objective of the present study was to design a well tolerated regimen, applicable to the vast majority of patients over 60 years with MCL, while improving the limited efficacy of R-CHOP immunochemotherapy. To achieve this goal, we have used two novel drugs, i.e. ofatumumab and bendamustine. Ofatumumab is a fully human monoclonal antibody targeting a unique epitope on the CD20 molecule: its superior complement-dependent cytotoxicity compared to rituximab may translate into longer duration of treatment response. Bendamustine is a unique agent with both nitrogen mustard group and a benzimidazole ring: when given to relapsed or refractory MCL patients in combination with rituximab, it has shown a very favourable toxicity profile, as well as considerable activity. Methods:Previously untreated patients with MCL, any stage (patients with disease limited only to the bone marrow were excluded), aged ≥ 60 years, were enrolled into a therapeutic protocol consisting of six cycles of immunochemotherapy as follows: ofatumumab 1000 mg i.v. on day 1 (300 mg only for the first cycle); bendamustine 120 mg/sm i.v. on day 2 and 3; dexamethasone 40 mg i.v. on day 1 to 4. Each cycle of therapy was administered every 21 days. Hematopoietic growth factors were used in case of neutropenia. Results:As of July 31, 2011, 36 of the 50 planned patients have been enrolled and 19 have received the whole treatment. Cyclin D1 immunohistochemistry was positive in 18 patients (95%), thus confirming the diagnosis, while in the patient found negative at cyclin D1 analysis, Sox11 was used to confirm MCL; 1 patient (5%) had blastoid histology. Median age was 69 years (range 60–81), stage at diagnosis was IV in 95% of patients; MIPI score was low in 8 (42%), intermediate in 8 (42%) and high in 3 patients (16%); there were 15 male (79%) and 4 female patients (21%). Bone marrow (BM) involvement was detected by immunohistochemistry in 15 (79%) patients and by polymerase chain reaction (PCR) in all patients. Involvement of peripheral blood was detected by flow-cytometry in 12 cases (63%), by PCR in 18 cases (94%), while no data were available in the remaining case. A molecular probe was available for all patients: bcl1/IGH rearrangement was detected by PCR in 11 patients (58%), and an IGH allele-specific oriented primer was designed for the remaining 8 patients (42%).Most adverse events (AEs) were mild or moderate in severity. The most common grade ≥ 3 treatment-related AE were neutropenia (10.5%), febrile neutropenia (10.5%), anemia (5.2%) and thrombocytopenia (5.2%). Two patients (10.5%) experienced tumor lysis syndrome: one was complicated by acute renal failure and the other by disseminated intravascular coagulation; both recovered with adequate therapy. Infectious complications consisted of one episode of pneumonia and one herpes zoster reactivation: both cases were resolved with antibiotic and antiviral therapy, respectively. Of note, 10 patients (52%) experienced cytomegalovirus reactivation, detected as pp65 antigen immunofluorescence, without evidence of infection: all these patients received anti-CMV therapy with negativization of the assay.Among the 19 patients evaluable for response, overall response rate was 94%, with CR in 17 patients, partial remission in one patient and progressive disease after the second cycle in one patient; 15 out of 15 patients evaluable for minimal residual disease in the BM showed molecular remission. Conclusions:Chemotherapy with bendamustine and ofatumumab appears generally safe and well tolerated to date in MCL patients aged ≥ 65 years requiring treatment. Preliminary data about efficacy are encouraging: accrual is ongoing for further evaluation. Disclosures:Off Label Use: ofatumumab, in label for CLL.